Historic, archived document Do not assume content reflects current scientific knowledge, policies, or practices. ages eee Issued November 23, 1908. U. S. DEPARTMENT OF AGRICULTURE. BUREAU OF CHEMISTRY—BULLETIN No. 115. “ H. W. WILEY, CHIEF OF BUREAU. “A PRELIMINARY STUDY OF THE EFFECTS OF COLD STORAGE ON EGGS, QUAIL, AND CHICKENS. By H. W. WILEY, Chief, Bureau of Chemistry, U. S. Department of Agriculture, WITH THE COLLABORATION OF M. E. PENNINGTON, G. W. STILES, Jr., B. J. HOWARD, AND F.C. COOK. rs ——¥$S COCA Ss = WASHINGTON: GOVERNMENT PRINTING OFFICE, 1908, bi Priel aN SMETTAL. U.S. DEPARTMENT OF AGRICULTURE, BUREAU OF CHEMISTRY, Washington, D. C., May 18, 1908. Sir: I submit herewith for your consideration and approval the manuscript of a preliminary report on the changes in food products kept in storage, especially at low temperatures. These investigations have been carried on in harmony with the will of Congress as spe- cifically expressed in the several appropriation bills of the last few years. The objects of the studies are fully stated in the introduction to the present report, now offered for publication as Bulletin 115 of the Bureau of Chemistry. Respectfully, EE: Wana, Chief of Bureau. Hon. JAMES WILSON, Secretary of Agriculture. CONE NAES,. Page. Bera amvOr Rem VestiGalOMe cul sss c eb. t cess e eee sete scores deeee "9 Information furnished by cold-storage warehousemen...:...............----- 1 SN IAOTD) COM UNG) Yue ase 3 Ne Sie ES len 25 |W ASIEG IS (ROT RE ES Cys) ee RU a SIR oh a 27 IP] GH GIR S ceo eee cice oo ad ei Ce Ee ae PH Fat sled Me NanreteMisplcc eae ees ees oa eo Lone ee ee 28 Ghemitcalanalysis:..222.2 2... - Eis 5 SHBG Stee Be ee ae ee 29 MISCuUSSTOMMOTC ata aes Rees ee Lo! Ee aI) Tk ag GE 29 CONGMISIONS Ma meee er sem ee Sete Pi a ad 30 PacreniolooicdleexamiumahlOMei 2. koe hse fo. sec ok nee oe ol 34 IROG DING WO acne oe SSO See ae Ie eco 34 1D FS CUBIS TOND. OVTRSETUI ESS Gk iS oc yc icc ee ee eg ec 34 Mierocco piealmexammlmaiionie rr hae aw te. woh eee Me ee 36 INGE BUS so BS Se oe cee SS eee Eel a eet ec 36 @aldestoracerecos mre se ee fee eee ee es Sa ee 36 Supplementary work on rosette crystals .-......-...-:--.----:-++-- 37 meeOuaticold-stored under knowm conditions....02 2.20220... 2022-62222 ee eee 37 DAAONS OUTS TANS SSa eo SAE & Be Reece ee ee er 37 acrenloloriealmexamminlattoneras 4-22.58 5 6 2 8-42 8 he bloc oe ae ee ticle = 40 mit. Chickens cold-stored under known conditions .......-....-........---.- 44 MroruGleneienestse sere sere ee ee a ne EN sh eek iwc 44 maeheriolocicamexamumatlOMer tase ee ek ok sw ee eae ee ee tet 51 ee viance: cold-storate chickens: 32.05... 2 2. es ol eo be lect eee oe kee 57 croc ChlonmeeuNnmnr De aM ee ee ene 57 Chemical studies........ dik OS cen aes eg SE ieee Ae die a ea 59 Résumé of the literature on the chemical composition of poultry. - - 59 Selechonyoi material sorstudy ern o.oo es. oe. hee be ee ee 61 Chemucaintechmioie = terre sh a) ck. fee ees nee 2 62 ibteparationoisample:t=: 2ecc-c\2.5..-5.~.. Ss Gee SOE eRe aes 62 Musclesdeternmmiations!s 2.22 292-5 .& oye. oes seks st ee 63 Mota lisoludsramGbwater eo 42 sk re ee ne ee ee ea 63 TRAM: setcomd © aes Oates aaa Oe eit eae ee Aa Se 63 IME 533 kod bke S 5.0 bee Os LEE eae eee ee 63 INDITOPEMOUS CONSHILMeMist. 8 ee ae ess ee ee eee os 64 ]DSG> CIGUCIATIVIBNTOSS SS actin Sa ee ae ee eee a 66 PAH SCSHOMIGES MCMC KONG 24 2 = aps ifac- s 2e deh weyse accesses es eee 67 Analyses olcold-stored-chickens ...0.:<../.-2.2-.1)-6222 8. seo 69 LS OSEAN CEN ST HOKG MESS, SS Gea eee a er 75 RCC MNCITC EMER) Spee nec re. Ol Ps fe Roce eos Saco hoes 75 Examination of fresh and cold-stored chickens .............------- 77 ASHG Here ee Mei T CSE eee ee a See SE oe ole on oe obo oe ow ee ee See be 79 sRGeericniem me teen ee tO ee So ie Gy See ee oe 79 Examination of fresh and cold-storage chicken muscle ........-.-.-- 81 6 PRELIMINARY: COLD STORAGE STUDIES. IV. Market cold-storage chickens—Continued. Page. General discussion 2222042 22. Gein San Oe 85 Resistance of bacteria to low temperatures __._:........-2.2=---- 85 Development of bacteria at low temperatures - _...-....------.--- 86 Inhibitory action of low temperatures on bacteria. ......-----.---- 88 Chemical changes produced by bacteria at iow temperatures -...--- 89 Effect of low temperatures on enzymes -.........--.------------ eS 89 The action on flesh of bacteria and enzymes at low temperatures .- - 91 Review of the literature =. = ss ee eee 99 Bacteriological and chemical investigations........- i ee eee 100, Molke eoso te Fee ee ee eee 100 Chickens! 2.222 a See ee ee eee 102 Histological studies -c2s2 58 Sees Se oe a ee eee 104 DDT Sa tc eee ei eM ea Spears eS aed Re Sef Sg Sis Sy ped ee) 104 Chicketis) 2325025... 3 AS A ee ee ee eee 105 Appendix 22.22.2257 ..0 6525 Se ls Sn ee eS Trend of legislation regulating the cold storage of foods, etc..............- 108 General discussion aroused by proposed Chicago ordinance..-...-.---- 108 Various opinions on drawn and undrawn poultry......---..--.------- 110 Sugpestive- legislative action 2. 228552...20232 2 ee eee 112 Massachusetts enactments - -2-- 6.52: 9. 222-3 ee eee 112 Sacramento; Cal. ordinance 228 4-2 ee 113 Knactments in Uhinois. 23. 22.2 Se se ee eee 113 Proposed Jegislation in New York 21:50) =. 2. eee ee 114 Proposed legislation in the District of Columbia-_.---..---.--.-.----- 114 Canadian -cold-storage act ..2 = 222 22 ae stone ee eee 114 Tata aie Ree ee PLATE I. Lt. EEL, IV. V. VI. Vil. VIII. — ra Ps s on XII: XT. . Soaked breast muscle of chicken in cold storage two years. . Breast muscle of chicken in cold storage four years. ILLUSTRATIONS. Fig. 1.—Outer shell membrane of fresh egg. membrane of fresh egg Fig. 2.—Inner shell Fig. 1.—Yellow yolk of fresh egg. Fig. 2.—White yolk of fresh egg . . Fig. 1.—Yellow yolk of fresh egg, hardened in situ. Fig. 2.— Rosette crystal found in yolk of cold-storage egg................- Chicken after storage at 13° F. from December 24, 1903, to February 6, 1908 Chicken, massaged and made as flexible as possible, without soaking, after storage at 13° F. for about four years Normal chicken muscle. Fig. 1.—Longitudinal section. Fig. 2.— Hliramsiviets CISC GhOlmE MEE mire ak ne awe em mew a ee SS Breast muscle of chicken in cold storage fourteen months. Fig. 1.— Longitudinal section. Fig. 2.—Transverse section......-.......- Thigh muscle from chicken in cold storage fourteen months, showing invasion of fiber by bacteria. “Fig. 1.—Longitudinal section. Fig. 2.—Transverse section . Unsoaked breast muscle of chicken in cold storage two years. Fig. 1.—Longitudinal section. Fig. 2.—Tyransverse section. ...... Fig. 1.— Fig. 2,—Transverse section........---.-.- Fig. 1.—Longi- tudinal section showing vacuolated degeneration. Fig. 2.—Longi- tudinal section showing varied and extensive degeneration....... Small intestine of fresh chicken Longitudinal section. Page. 30 34 36 0 12 80 82 82 84 84 84 86 86 ie te mt he A PRELIMINARY STUDY OF THE EFFECTS OF COLD STORAGE ON EGGS, QUAIL, AND CHICKENS. GENERAL PLAN OF THE INVESTIGATION. In the act making appropriations for the Department of Agricul- ture for the fiscal year ended June 30, 1905, Congress authorized the Secretary of Agriculture, through the Bureau of Chemistry, “‘to inves- tigate the adulteration of foods, condiments, beverages, and drugs, when deemed by the Secretary of Agriculture advisable, and to pub- lish the results of such investigations when thought advisable, and also the effect of cold storage upon the healthfulness of foods.” In order to carry into effect this provision of the act of Congress, a study of the effects of cold storage was organized with the following points in view: First. To ascertain in so far as possible the kinds and character of food products kept in cold storage. Second. To ascertain the minimum, maximum, and average length of time which such products were kept in cold storage. Third. To ascertain the usual temperatures at which foods were held in cold storage. (To secure an expression from the trade on these points a circular letter was prepared and forwarded to the leading cold-storage firms whose addresses could be obtained, accompanied by a blank which the persons receiving it were requested to fill out in as great detail as possible. Copies of this circular and the information obtained in reply thereto are given on pages 11 to 24.) Fourth. To ascertain the effects of cold storage upon the organo- leptic, chemical, bacteriological, and histological properties of stored foods. In planning the different lines of investigation which should be car- ried on the following points were considered: It is evident that the effect of cold-storage must be in the first place to inhibit, to a very large extent, the ordinary processes of fer- mentation and decay. It is well known that bodies subjected to a temperature below that of the freezing point respond very slowly and very incompletely to the action of the ordinary ferments, so that practically it may be stated that, so far as actual decay is con- 9 10 PRELIMINARY COLD STORAGE STUDIES. cerned, the tissues are almost as well preserved as if they had been completely sterilized and subsequently protected from any external — infection. That changes, however, do go on, even in frozen bodies, is incontestable. The nature of these changes, the method by which they are effected, and the results produced are, therefore, all legiti- mate fields of inquiry in attempting to carry out the will of Congress. It is evident, then, that the changes which take place are of the fol- lowing character: First. There may be bacterial or enzymic action taking place in a very limited degree and yet of sufficient magnitude in the course of time to produce distinct results. Therefore, one line of investigation - must evidently be directed to the ascertainment of any changes of this character. Second. The activity of the organisms above mentioned must necessarily produce changes in chemical constitution which may affect very markedly the nutritive properties and, to that extent, the wholesomeness of the food products. Careful chemical examina- tions of the food products in a fresh state as they are entered at the cold-storage warehouses and also a similar examination on their withdrawal at different intervals are, therefore, necessary. The object of these examinations is, of course, to determine the nature and extent of the chemical changes produced. Third. From the action of the above agencies it is evident also that changes in the actual construction of the tissues may be expected. These changes, which are known scientifically as histological changes, are not evident in their incipient state to the senses, and therefore can only be detected by careful microscopic examination. Special microscopical studies have therefore been made in the investigations, an account of which is to follow. - Fourth. The effects of storing certain food products, such as fowls and game birds, without removing the contents of the body cavity, as compared with changes which go on after removing such waste material, present a problem of great interest in connection with the examination. To this end, in the case of fowls and game birds, examinations were made of those stored after drawing (which is the technical term used for the removal of the entrails), and also of those stored without drawing. Birds of as nearly as possible the same age and in the same condition were stored in both the drawn and the undrawn state and removed from storage from time to time for examination. : Fifth. Among the most important of the changes which may take place in cold storage are those which relate to organoleptic properties of the foods. The color, odor, taste, tenderness, etc., of these food products have much to do with their value as food and with their effect upon health and digestion. To this end careful studies were DATA FURNISHED BY WAREHOUSEMEN. ea inaugurated relating to-all of these organoleptic properties. Juries were constituted to examine food products when they were placed in cold storage, and at stated intervals thereafter, for the purpose of determining the character of the changes which may have taken place in these properties. i Sufficient progress has now been made in the work to clearly out- line the exact nature of the problems which are presented, and to war- rant the publication of the preliminary report. The problem, how- ever, is one of such magnitude, and one which requires so great a length of time to study all of the conditions which have to be taken into consideration, that it can only be said at the present moment that the work has been fully inaugurated and many of the difficulties which at first presented themselves have been surmounted. A rec- ord of about two years of continued investigation has disclosed a suf- ficient number of valuable data to warrant their publication, not for the purpose of drawing a final conclusion respecting all the questions which have been raised, but rather for placing the matter in such a light as to clearly indicate the character and magnitude of the work still to be done. INFORMATION FURNISHED BY COLD-STORAGE WAREHOUSEMEN. As was stated in giving the plan of the investigation, a letter asking for the cooperation of the cold-storage warehousemen was sent out, together with a form covering the points in regard to which informa- tion was desired. The response was most satisfactory, showing every indication on the part of the trade to cooperate in the investigation, and the correspondence in condensed form is submitted as containing valuable material for comparison and study. The letter sent out by the Department read as follows: Avueust 15, 1904. GENTLEMEN: An act of Congress making appropriations for the Department of Agri- culture, approved April 23, 1904, authorizes the Secretary of Agriculture ‘‘to investi- gate the effect of cold storage upon the healthfulness of foods.’? This is an investiga- tion in which the producers of foods, those who keep them in cold storage, and those who finally consume them are all equally interested. The wholesomeness of foods is a condition which depends on a number of factors, such as soundness, freedom from products of decay, proper balance of food elements, palatability, etc. It is well known that cold storage, at a proper temperature, improves certain food products up to a certain point. Such is the case with fresh meat, poultry, fruits, etc. There are other food products which it is well known are not improved by keeping in cold storage at any temperature, such as fish, oysters, eggs, etc. The exigencies of transportation and market conditions, however, often require that such food be kept for a time before consumption, and cold storage offers the best means of protection in such cases. It is important to know how long bodies of this kind can be kept in cold storage without materially lessening their value as food or impairing to any appreciable extent their wholesomeness. For bodies of the first class, which are improved upon _ keeping, there must be some definite period of storage which develops their maximum qualities, both of palatability and wholesomeness. 12 PRELIMINARY COLD STORAGE STUDIES. In carrying out the will of Congress, I desire to secure the collaboration of those engaged in the cold-storage business. It is important in the beginning of the work and in the preparation for carrying it on in detail that the general practices in regard to the period of detention for various forms of food should be known. I have therefore submitted for your consideration a blank requesting certain infor- mation which it is hoped you can give without in any way revealing trade secrets that are regarded as private property. In answering the questions, which I hope you will do at your earliest convenience, I shall esteem it a favor if you will suggest any lines of investigation which in your opinion may be helpful in attaining the object that Congress had in view when author- izing this investigation. All correspondence on the subject should be addressed to the Chief of the Bureau of Chemistry, in which Bureau the investigation will be conducted. Respectiully, Wis L. Moore, Acting Secretary. Inclosures: Blank form for information. Return addressed and franked envelope. A copy of the blank accompanying this circular, as filled out by the commissary officer of the U. S. Military Academy at West Point, is given below: Foop Propwucts In CoLp STORAGE. 1. Name and location of company: Cadet Mess, U. 8. Military Academy, West Point, NETS 2. Principal food products kept in storage: Fresh meats of all kinds, poultry, fruit, vegetables, butter, lard, all the usual perishable food products required by a first- class educational institution. ANIMAL PRODUCTS. 3. Fresh meats (fresh when stored). (a) Minimum time of storage, 2 or 3 days. (b) Maximum time of storage, 1 year. (c) Usual time of storage, 1 to 3 months’ supply of beef purchased at a time. (d) Temperature of storage: Meats kept at 30° F., except one lot 16 hindquarters beef kept at 15° F. for experimental purposes. 4. Preserved meats (hams, shoulders, sausages, etc.). (a) Minimum time of storage, a week or two. (b) Maximum time of storage, 4 or 5 months. (c) Usual time of storage, 1 month. (d) Temperature of storage, 30° F. 5. Eggs. (a) Minimum time of storage: Used immediately to 1 week. (b) Maximum time of storage, 1 month. (c) Usual time of storage, 2 weeks. ; (d) Temperature of storage, 34° F. 6. Fish (fresh when stored). (a) Minimum time of storage: Not stored usually. Used immediately when received, though it is sometimes necessary to hold in refrigerator from 24 to 48 hours, which is done without any ill effects. Occasionally a few fish are left over from the lot and are put in a cooler where the temperature is at 15° F. These small lots have been kept in this temperature for 6 months without in any way deteriorating. DATA FURNISHED BY WAREHOUSEMEN. 1.5) 7. Oysters (fresh when stored). (a) Minimum time of storage: Not stored longer than 24 to 36 hours. (b) Maximum time of storage: —— (c) Usual time of storage: —— (d) Temperature of storage: —— 8. Poultry (fresh when stored). (A) Drawn. (a) Minimum time of storage, 1 week. (b) Maximum time of storage, 3 months. (c) Usual time of storage, 1 month. (d) Temperature of storage, 15° F. (B) Undrawn. (a) Minimum time of storage, 1 week. (b) Maximum time of storage, 3 months. (c) Usual time of storage, 1 month. (d) Temperature of storage, 15° F. (C) Is poultry in your house usually drawn or undrawn? Undrawn. 9. Game (birds). (a, b, c) Minimum time of storage: Experimental lot of 12 pairs fresh-killed partridges put in storage November, 1903. One pair used every 2 months. No deterioration noticed. Last pair now in storage appears to be all right. (d) Temperature of storage, 15° F. (e) Are they usually drawn or undrawn? Undrawn. 10. Butter. (a) Minimum time of storage, 1 week. (b) Maximum time of storage, 1 month. (c) Usual time of storage, 2 weeks. (d) Temperature of storage, 34° F. 11. Cheese. (a) Minimum time of storage, 1 week. (b) Maximum time of storage, 3 months. (c) Usual time of storage, 6 weeks. (d) Temperature of storage, 34° F. PLANT PRODUCTS. 12. Apples and pears (summer and winter). (a, b, c) Minimum time of storage: Small quantities of apples and pears for immediate use occasionally stored. No effect noticed on apples when kept two weeks. Pears, however, put in green ripened in 1 week to 10 days. (d) Temperature of storage, 34° F. 13. Peaches. (a, b, c) Minimum time of storage: Not stored, except small lots for immediate use. Have kept a week, show signs of decay, and then go very fast. (d) Temperature of storage, 34° F. 14. Strawberries, raspberries, blackberries. (a, b, c) Minimum time of storage: Small lots for immediate consumption stored from time to time. Decay very fast in spite of refrigeration. (d) Temperature of storage, 34° F. 15. Grapes. (a, b, ce) Minimum time of storage: Fifty crates selected Concord grapes, all per- fect bunches, no overripe, immature, or bruised fruit left on the bunches. They became musty and dried, falling off the stems; experiment unsuccess- ful. Were stored 2 months. (d) Temperature of storage, 34° F, 14 PRELIMINARY COLD STORAGE STUDIES. 16. Oranges and lemons. (a,b, c,d) Minimum time of storage: Oranges have been stored for 1 month and lemons have been kept 3 months with good results, when the fruit was per- fectly sound and dry when stored. When wet or decayed when stored, cold storage did not prevent the spread of further decay. 17. Green peas and beans. Not stored. 18. Tomatoes. (a) Minimum time of storage, a few days. (b) Maximum time of storage, 2 weeks. (c) Usual time of storage, 1 week. Very successful with these. (d) Temperature of storage, 34° F. 19. Cucumbers. (a) Minimum time of storage, 1 week. (b) Maximum time of storage, | month. (c) Usual time of storage, 2 weeks. Very successful with these. (d) Temperature of storage, 34° F. 20. Celery. (a) Minimum time of storage, a few days. (b) Maximum time of storage, 3 weeks. (c) Usual time of storage, 1 week. (d) Temperature of storage, 34° F. Very successful with this. 21. Cabbage. Not stored. 22. Potatoes. Not stored. MISCELLANEOUS PRODUCTS. [No statements. ] A large number of replies was received in response to this circular, and the information given has been compiled in tabular form as follows: Tabulation of data furnished by warehousemen. (A) ANIMAL PRODUCTS. [Representing 43 firms.] FRESH MEATS. Number of firms || Number of firms Length of time in storage. averaging this | Temperature at which held. averaging this | time: temperature. Weeks. OU Ones 8 sears cacti eee Two. ll =B Glows O PLR tees sees oars nee Three. ATWO eG arse ates a ah ees eee Seven. LIS SOrGOp Oss ae eee es i ene Twelve. PPT CG Seas see ech d pase a eee One. NOM O24 ses ee eae eae Ten. A DOU Be) eee ee et en NS a Six. 25: tO: Sleek ese ee ee Hight. ES) Bee ee Sy ay a ae ag a EWOw es S2COVAO ee ee ee Six. IOAN oe ois Gace eat te eal eR Four. Over ciehits 33s eee Twelve. Average: maxim times e275 Fe ee Se Se a eee ct co ap eee ce days.. 201 SA VOL Agee TAIN TT WI CLIVE Leaps ye ae EIT aay eh ee dokes rely, FAVELA USUI Al: Gili e ey ee ites MeN ee See ete acre Dao Dee SONS 5, Se ey a cy GR Goze eeaoS Average temperatures sos scissor eS ee es a gna ea oes eae OSS Ws) RemArKS.—In many cases the firms differentiate two classes of fresh stored meats, namely, frozen and unfrozen. The first named is kept at lower temperatures (—22° to 10°) and for longer periods of time than the unfrozen (kept at 28° to 40° F.). DATA FURNISHED BY WAREHOUSEMEN. 15 Tabulation of data furnished by warehousemen—Continued. (A) ANIMAL PRODUCTS—Continued. PRESERVED MEATS. | Number of firms Number of firms Length of time in storage. averaging this || Temperature at which held. averaging this time. temperature. Months. mt. | MOMs ste ateihemniee nero hs ae'ss Seven BOLO Wal Sem pecs ne ey win oe oe RE | One NOP mek en nn aie ccd Some Three OFC ORION eee rea Rites | Two MCCS et a sas 2 oa io ake Golsne Nine. ORG Op 2A Se ees ee eee Coe. OUR EGS See ee Smee nee memes Two. AUG ORo lee ee ec cee ee ee eee | Seven. Veer e eter. aries Becca ele One. DA LOVAQ Baas ee eine ce es teehee | Fifteen. SE. 6 eae es er ae eS ae Two. OWerrAQur ec ecco sae ames ee One PIOMOM eee secre Wao Nee. ee monton sg iD TEAS SAE Op Sey eee ets a eee ne Two | OLAS CA) AL CMNULIG NIU Cra eters ee cee cei Me Ae eens. oboe ceils oe tiee weeee ieee days.. 193 PC OR A SOMME DRI Cae meat erat ae cre Sere Nera har Note ol Gi eee cece Sreccin rs tera ciel ate wiatele ete nee bie he does) 34 WM EGECATSER I HRTIBS Seco Sent eae ite COS Cee Se Io JE Sen ee gags eee ee dome 84 JASHGTEE@) HOLT OSE AUERE Bo co apocrine tic eet Taio aiee Gee eee eter eee ee ee Ae 28) REMARKS.—In many cases the firms differentiate two classes of preserved stored _meats, cured and uncured, and prescribe different conditions for their storage. Under the seven firms for whom one month is stated as the average time held are included those giving shorter periods of time. This reduces the average, which is really just ‘three months. EGGS. Number of firms | Number of firms Length of time in storage. averaging this || Temperature at which held. averaging this : time. temperature. Months. | OA OM@snd koeasaaede aes pepse ese Six Glows 0 Sela hee see note eceine TNWO).s3 GH ae See Eee eae ee One ORC OS Oe ree se le. les One TRS Soe So ae ee ee Five ORDO 2as BAe etm ae tee ary POU S308 Gaoaae sees aeeeee DAG ORS le eee ess st errean ee Twenty-three. INN @s5 4 See 5 ae eee ee Three. SOREOPA (MR acy ry Swit Ere tee Sk Eleven. STG AIES Circe SoS pC eee Eleven. Onens4 02a ees Seen ce Set SiOVGIN 2 eee ae le em ee eee ae Four. EO SUD ie re Ce aes en Re ree el Seven AGUS MOS Sh aM MAF BOAT LEY So Asan Oe easement SR A eGR fra a es SOU i ear days.. 256 Psa O CRIMI UNITING IT Ota Marcie e esi ey Sm tap.s seekers eet og. Dee at ee Bree eee ne MMAR Ml meee Gomer e449 BrAsverace usual time... 355... ee tens. Se eee ae ee ea: ROM pel ate as ae, A ve Sos Bs aye Ae CS GOseee eb ASTGRDISE: GL OOO e A LEG oy ae GOSES Eo Bees SOB OOo SOOEE 6 COR Ree ss Sree ness ee eae ae SoS OSS REMARKS.—Under the six firms for which one month is stated as the average time held are included shorter periods of time, reducing the average time, which is really about six months. FISH. : : Number of firms Number of firms Length of time in storage. | averaging this Temperature at which held. averaging this time. temperature. Months 7s 2 TG SE Re ae eee | Two. Belowaleecme sie cece san foe One. SP Cape ee ea Pe et | Four. Oup OnE ORR eae os eee Min Eight. “TEI RG 2) CS ASR ReaD rs lee ie ee Three. NOS CORZA Se ie ene ie ie Thirteen. CUE SS Me ee | Four. DDLUOR Se isin echt Saas trols 25 One. LID AGE Se ae peal See eae Three. SOA ® eerie cake soot ee One SU Neem tavs s SUS a Se Pe ar Bo Six. Owen 40h: nae 22ers wee see ee SEVOD SA SRR ee Spee ete 12 TaN 5 Sak eae ieee caer eee One AP OPNGS ene parianbiaa Tea SS GM Sa 5 rs ms i Nem SS greys rn days.. 229 MRE MCHSC ARI SIRT IE GATTO oe ee Meme PME. aah A! ney a og n e B Oa ee ey do. 34 meet UA U Eee Seen ee ee ee Me eta i ee Ce do 133 J SHURE) UGTA OSEAN SES, aes one a PN BY 00 RS DO es oie eben ue na Olea cs deh tv ery Caltrans ON we _ RemaArRKS.—Several firms state that they freeze the ‘fish at once at temperatures from —15° F. to 5° F., then carry at the temperatures given. 16 PRELIMINARY COLD STORAGE STUDIES. Tabulation of data furnished by warehousemen—Continued. (A) ANIMAL PRODUCTS—Continued. OYSTERS. | Number of firms | | Number of firms Length of time in storage. averaging this || Temperature at which held. | averaging this time. | temperature. , | Weeks. mR One? [52k ee eee | Four. (Below: 022222 522 see ere TT WOS 2 S222 Sa eee ee | Three. OO R Qe eh ee ae eae | One. IBN Tee setae ee eee nes | HOSE OS24 ase ce ee ree One. . HOUR SS eho biel Nn See kes 72559 HO Leen ls ae a, Three. . IVS ea ee ea ee eos SU bOPAOS : teeta ces San ee eats Seven. | Sixes S203 Saag Ne cee ee | Ower 402 e be ne re ee see S@VenS o> 22 55 ae een eee igh Ska oS Berne ace eee One | | More thanieight.2 2-22.55 One | | > Average maximum Gime. 22%. ee ee ee days.. 50 /AVELAZe MINIMUM ELME 2 2S 2a ese CS a ee agape eae lS eee dosta ts AVErAge Usual CiMe. = 25 eA 2 a Se Rt pe eR Ee ee ea ane ee dos 229 Average temperature: js. 6 5.5 S25 5S a es a I a ee fs Cr a DRS 786 ReMaRKS.—One Alaska firm keeps oysters a much longer period than any firm in this country. POULTRY (DRAWN). | Number of firms Number of! firms Length of time in storage. averaging this | Temperature at which held. averaging this time. temperature. Months. ae ie Onee eee ee a ae Two. Below. O02. aseis- sso te ee eee One LE Oe ne eye rye ee Four. [le 1OSGOre Os ee eee Seven PIT CS sneer ee ele eae, eae Four. el OSGO]24 Sat ree eh re ee ees Seven HOUT a= Sane eae eo ee pe ee Three. | e256O SL eae x oe ae ee eee Two PVE re te en a eee ee Two. [|:32 COAOl Sa 5.c oes. Se ees Two Sik ee coe oe Se ae eae One ROVER AQ: = Aaa 2 so ee aoe SOVGT Ge cere ace nh ee coer A Ger Sin, oho ea ne More thanerzh tas= 5" =ese ee One AVETALS MAXIMUM TMC 25s oS See ye epee am NS 2 pee gem days.. 219 AVeTAge MINIMUNY TIME = 222s oe et ee ee Ae er ee eee a Gols ee 34 A VOTALS USUAL AGING a seo ee se ae aa ya ey as ea dose ett2 ‘Average’ Temperature soe oi ok Sy Se SE Pe ie ae ane ee nan ne SRS s12:4 REMARKS.—Seyeral firms who handle undrawn poultry state that they do not handle drawn poultry, and one firm says experience has taught them that the undrawn keeps better than the drawn. : POULTRY (UNDRAWN). | | Number of firms | Number of firms Length of time in storage. averaging this || Temperature at which held. averaging this | time _ temperature. | | Months. se On Fe eae te eae Two Below 0 ae 5- 455 Sa ae Four PW eS cnet es Four | XO RG OO ers ee ee eee eee Nine iPhree sass A. See ee se Soest Three || AONGO 242 ee es See ores Sixteen INOUE a= aa en ere | Nine heen oe bee See creer tS TE Two IWiVeG2E "Soo ea eee See ae | Four. We S32 EOrAOs Oe ey ae are ere eee Two SEX. se aaa een ee aes ee | OUI | ONCE: 40 Re eee SC Velo see a oa see eee One. Hight: S22 Se ae eee One More thanieishGas ae eee | Two Average maximum time: gos. 232 Sao So Ss ee Se re ae ee ee eee days.. 256 Average minimum time ~ 2/22 2..=e- oe a a a eer Ste ee ee dos ae S2 Average Usual time 2. 2-2 cc See ee ee re eee doe sIst Average tempera tuless2- 2-5 eee ee MGseb Sock eee Se ee ee SRA TIS7 ReMaARKS.—In answer to the question: Is the poultry in your house usually drawn or undrawn, seven firms repled ‘‘drawn” and twenty-six ‘‘undrawn.’’ Sixteen of the latter stated that it was always undrawn. Three answered that they stored both, without stating any preference. DATA FURNISHED BY WAREHOUSEMEN. lil Tabulation of data furnished by warehousemen—Continued. (A) ANIMAL PRODUCTS—Continued. GAME (BIRDS). Number of firms || Number of firms Length of time in storage. averaging this || Temperature at which held. averaging this time. temperature. Months. S406 ‘OWOSYS5 CRedboae sdocee Cree ae One. BOLO Ob acta cecal festa One. IN @s She aRe Seek ees Bae oem ease ae Six. OFCOM ORE ce ee eee eee cee Six. BMINO OM ery crates cae noe oeraye claret sins | Two. OW OLE eS aein se icc Se eene cere Thirteen. [ROUTES 3 Sates ape renee eee ern Four. DASA FO) Gl ee nee ee acne eae ne ee Two. WIV Gis neato SB ee eae eae eee Three. Dob OLA as seer Ses eee ars ral rete Two STi Sa SRG eee eee ean OM CRAQ RE Mea tee ia eens SENG. oB ae Eee ee ESBS ee Ree ens IDVENE Be Gata eee ene ee one More than-erghit. 2. 2222--.5-- Four. JASN ED Ta ab oo Sboay ities A soe eee ae ee ee age Sold ee ee ae gee eee eens days.. 275 SCOT) ORIENT NULTAN GCI) © Retr ee pattem ae eT ey BNE Ee UN ae Ae Sk cele’ seinae webs do. 29 J SIGTRAED WET WSs 3 boS as Seitole, dc es ROE ree Se I Oe RETR ere er rear eae ea do. 133 ASCHER EG) (HDT SVR SS eras oe eB a Ge ee AP I te ae Ne eee “de5 iG REMARKS.—Two firms stated that they were controlled by the game laws as to time of storage. One stated that all game stored should be drawn, since it is shot. In reply to the question whether they were usually drawn or undrawn, three firms stated that they were usually drawn, and /ifteen, usually or always undrawn. One firm said they stored both kinds, equally. BUTTER. Number of firms | Number of firms Length of time in storage. averaging this | Temperature at which held. averaging this time. | temperature. Months. DIR, (ONG 5 5 Sees ee en ee eee ee Two Below AO eee eee eee Eight. TNO SGb See eee a are ere One. OF CO OUR wee nas cee Fourteen. UN GREYS “Sm AES OS ae ae et a Three ORC OS24 ae ae cee ene Thirteen. LOU sa ee Sm teem aoe Five ES TROUBI LS ss rea ears Sema eee eee Three. TPIN@) s cote ees ee ee Seven DOE OVA ai Se Meeps Heer teak oO Two. SESS us 5A eee eee Twelve Over QR ae eee es SCWEM SEAS ee er a ae eee Three IDTEIIRE 2 OG eee ees ae ee aes Five Morenhan elehibec.s92-. 02.0527 JA VCTRELE) TAG aha CRIO | BOTS 5 cele sea RRS IS NCR eS Se ae aS er re ee SP days.. 301 SVORDED miviarhmnonn (nb) SG Spb eebecose ae Bose 5 Dooce COLE CES Beno ace ee OOO eRe Smee Seige seis do 39 NCCT ES) TASTED THRIVE Rem ee ARIE eee a a do. 156 AICHIDEDS UH COMMIS boos sede dds SASS Ee DESEO Cee Oe te eS OSES Se CEI eat Chel eae een ene mis. 288 CHEESE. Number of firms Number of firms Length of time in storage. averaging this || Temperature at which held. averaging this time. temperature. Months. 1 TDL 2 Sa es ee Se One Bellows Se etee scene eee UNWO SG Sere see eee re eres see ee One. OCOMON aa oo Sees ceases One. DIED: Go eee Gates eee eee are Five ORL OR2S Oe etteee sees chee seas Two LOUIE SS ee ee angen en eae Six. DONS Lec cleo nesemes sae cee Six. IPI Qc c5 te SOO ee ee ener Seven So GO AO Nt iecis eane Soe eee ee Twenty-three. SIRS. GH SEE ee eee ae Nine OverA0n eee Sra jedi pees SOVGIDG Gt Eee ee eet Four [EAE 2 eek a a One. More than eight ............-.. One. ODS Se TTS CHOTA) PERSE Se ace ee oe en ee eee eRe ee ee days.. 292 ERROL SA Cet ll MIM URTTIROLITIO mentee ee eae eter ee pe ee SS ee ae tae Bhcee do. 42 A CTEREG) STOUT GLE eS SS A ES ek Pe Oar eee do 146 ASIST) PENG TOE TING = os totes sas ee bot qe a eke tee ane ee Oe oo eee ee ee ANS BAH 49078—Bull. 115—08——2 18 Tabulation of data furnished by warehousemen—Continued. PRELIMINARY COLD STORAGE STUDIES. (B) VEGETABLE PRODUCTS. [Representing 43 firms.] APPLES AND PEARS. Number of firms | Number of firms Length of time in storage. averaging this || Temperature at which held. averaging this time. temperature. Months. =O Ome ere ree ne tee Rema Five. Below; 02a ener ee ee DEW O22 soon ete Sel ang Three O° COn9 Be eet sae sae eee One. Mnreess a ae ee ae eee Four LO} GONQA GE ee Me ee ee ie ie ee OUD Rarsce ee eee ee Eleven ZDECOy SS ee eee Thirteen. BEY Cin en ees Se ee eS Eight 2) COS Lae ee ae ete Se Eighteen pS) Diane arate ote ene ae ON reat ei era Six. SO COVAO eee Sal ee eee Four. S@vi@n Saye ae ee ea ae pete One. ONGIIAQE 3a ae Re re eee | Ded al hegenenr spear hee al oe ea eRe Se ee More ithanrveight 2522-5 - = AGEL AGC TTA RATMULI (GLITNC see es oe ee Boe SR Ss tA re a SR Ee eee days.. 188 ASV OLAS TIVELY NUIT GLI ee I I a eee coy Se Cp eee ae ee do 34 Average USWA sblMe se. s2 236 CUCUMBERS. | Number of firms | Number of firms Length oi time in storage. | averaging this | Temperature at whichheld. | averaging this j} time. temperature. Weeks. | 1 °F. OME j= oo eae ee Se ees | Five. Below 0. 5-4-5522 wos 45252 eee ee ee Four. 1! 060, OS Sao ie Se eee SPR TGG Ee he ea eee ane Ras NOSCO}24 2s ee el ROUES A2 oot eae ee One. Dy COS Ess Lee ee ee ree ae One. RIVE Soha aoe oe ee eee [chee DOGS eye ae Seen een pee Three OPS ae oe eee ee eee ee LSabOAQs 250 sen eee oe eee Nine DeVere 2a ee Over 405. 2380 8. ees DY (23 0 Roe a es ree oe RS More than eight-<. = Average Maximum ‘time: 2... 5.52 es oe ae a ee days... . 36 Avetace MmimMumM | fMe ss. 232s SF 8 ee ee ee Sdo3e2 5 ‘Averagse tisual time 5282 2 S285. Ses ee ee dasa 8 Average temperature... 20 22 So -3 5: 53 so ee ete 28 CELERY. | Number of firms Number of firms Length of time in storage. averaging this || Temperature at which held. averaging this time. | temperature. Weeks | Coe: One oe yo a6 ee ee ee || ‘Belowi0s_..- 222-2 eee IP WOe oe eee a ae | Four 0609 22 ee SPATS 69 ees eee ere Four | LObo'24265 23-2 5s ee OUTS ee ae eee Five. 25 tO Bisce ss eee eee EVES ase ee ee One. | RBZ Gs ee eee ee Twelve SEX Feb cae eae eee eaesey One. 30.60 40522. too oak eee Eight Sevens = 22545- es at | Over40 2s 5 Si be esc ee igh G 22-2 22 eee ee eee Three More than eight=) = ==. 32 One. Average Maximum thine ss 3 = ee eee Pe ay Py RN cg Ee days... 71 Average minimum times. 22 29. 20 5 ss ae ae ee doz awit Average Usual time >... = 225 25 Soe ee es ee eee ee ee d0s=5. a0 Average temperature 2... sec coos e cas de aas oe nesne oe a see ad ee Ge ee eee SRS S454 DATA FURNISHED BY WAREHOUSEMEN,. 21 Tabulation of data furnished by warehousemen—Continued. (B) VEGETABLE PRODUCTS—Continued. CABBAGE. | Number of firms Number of firms Length of time in storage. averaging this || Temperature at which held. | averaging this time. | temperature. Weeks Cone | ROTM Ae cjsmle aie Oren nin Secale IBS CLOW O Rear ei erelaureatine hee ae | IMWOc's ge Bae pGe eee eee One. OREO 9) a re a es eet | One TIRED 2 ce eer er a oe Two. ORG ORI Ate Nae te ee teers SOUR Tete een ed ot ates cies Ay ai Two ZouT OR as Aneel er na arcie en rsa on BVO Nee eos eee: Soe So ti One. BUIROGY ooh BaD nereoSrE aaa ees Ten SIRS Leg B ep ReOe ewes Oo emer ee Two. SEMUKO) CLO ree et iene ae ane ae re Four SHE GUT a See os Ee eae OV ETHAN ie oto esae ma aeoe a | DTU s ca cubes ae So pepe ao ee aes | Three More than eight. 22... 25. 5..-2- Five JATORSG Tine chan ran Wt eae LSS eoe uel e eS SROs Hp OoUnE He One ES pea E Eee Gece neeE ements days.. 96 AVOTRTEA Tao DIAN THON. GES Bese eee e CoB OO ON CBOE COO BEC eee Se eee Bea ee nee See renee ae Gorse 24 AWD ES EWEN Tb) Bas Be ae SH a Ae Be ere eee ies nen aI Me she Ieee cee SNe a OOsos. BY METAS CHLC MMOL LLC weer sae arene eae oe moras ah ce atehe Soe ye eI oe Se ake Miulare soe Sislele etaene Sus No 3 BLA POTATOES. Length of time in storage. Months. Average maximum time Average minimum time Average usual time PMBGEN ACRbOUNDC EA GUNG wee rrtart etek may eer ai gerd tA to Seip tele a G Ren curds wae Sh acinar tare cekelere ole Length of time in storage. Number of firms | averaging this time. Temperature at which held. (C) MISCELLANEOUS. CONDENSED MILK. Number of firms averaging this time. Two. Two. Average maximum time Average minimum time Average usual time Average temperature Temperature at which held. | Number of firms averaging this temperature. Six. Eleven. days.. 133 CDscco Ae CMeos0 Cl NG mt) Number of firms averaging this temperature. Two Two days.. 165 dose 40 domes a el23 K°.. 32.3 92 PRELIMINARY COLD STORAGE STUDIES. Tabulation of data furnished by warehousemen—Continued. (C) MISCELLANEOUS—Continued. NUTS. | Number of firms || Number of firms Length of time in storage. | averaging this || Temperature at which held. averaging this time. temperature. Months. ska | °F, ONnee ee Se en ee ee | Below 0.225 ere ere EW cue ee ere ne O'CO;2 ee ea ee ee IDA TEGO: Bees ss aes Sh ie nee! | 10 GOD4 seco o aera nny eee HOUR TESS oot ee ae eee | One. D5 CO SUS ere 2 eee Fee eee One VO S a2 os aoe a eee One. 32). 60)402 Soe es aa ee Two Sixt eet ee to eee me ne | One. — OVER4A0 See eae ee eee BG VCMT se een ere ee High Gee Sa eee eae eee More) thanieights. 2225-02 eee One | ASVOTASE TIAA MUU SEITE es SSS are tee ee ate tee ee rc Sa pete oar en Se eee days.. 180 ANGLE LE MINIMUM SEINE se ooo so ae es Rei ee ee ne ee ee GOzg 23 AVCTASE USUAL: GING Sys Ys a SS Ae Se EN ee re at a ye re GOses= 155 Average temperature: a2 55 Sis hrs os Soca a ha ES Se eo SS Se ee ee ORs 3ls6 MOLASSES. | Number of firms | | Number of firms Length of time in storage. averaging this | | Temperature at which held. | averaging this time. temperature. Months. | . | oF, OneseeR era se ee MB Clow, Osseo ee ee SSW O ee ieee a ees eee ete b O'tOW9 See Soe Sener AMS aN Ree eee ea oe ei eee TOROS 24 Sea One ae ee nee FE OUT ae tere rc ee ed One. PA RO ed haem Shee Sak 3S 1 DIRS Beat ee eee ee ath sae S260) 40 are cere ore | Two. SS cera eae eo ey 8 eS Layer eae | One. Over40 Si ee eee CV EM ne Ae ets ak See co Ba SiGe ees Ge ee eR aia ty ae ae es More thanieiohite se. a... 2-55 Two Average Maximum times: 5. 2 sea oe LSA SS as os a ae On ee days... oe Average TULLE TOUT EA TIN a a aah Ie a aaa a ae do Average Usual times: 25.2 h oes ene es eet ee eee do 150 AVerare Temperature sss FF Ae es et Sl eA a aa ear oF 34.5 DRIED FRUITS. Number of firms | Number of firms Length of time in storage. | averaging this Temperature at which held. | averaging this | time. temperature. Months. | | oF. Oe iee ss ae e ee Bete ses aay || Below Oc ee oe eee AIWO Remi pnt were ees Aas co beak Ue roe? etree ed pe Oe MNT eter Senet es eek eee | One || tO Of 24 Rees een TROUT gates yh ae ee | Two 125 tO; 3) 2252 sae eee One [DID Ope eenie cease aueEs San teens One NBG YA RONG Y Sees Sear wom ee | Three eaten oo oh a Se heen e eae | CO |: SoxCO! 402 Se Soo eas eee _ Two SOV CME ee es Se Super eae MOC 40 Seo eae en | Dited oN uae ee ete ee ape ele eer | More than eight............-_. | i} A VELOSO Ta RUMMY, CATO eas ee ae ae ye days... 180 Average Minimum Me SS Ss Se a a Sp es Re eee do: esa 30 ALVEOLAR SS Ustream do---- 140 Average temperature 2.25.22 22 ye ee ee ee Op 39953 : PRODUCTS REPORTED BY ONE FIRM ONLY. Rabbits: i Minimum time of storage, 1 week. Maximum time of storage, 2 months. Usual time of storage, 2 weeks. Temperature of storage, short storage, 32° F.; long, 17° F. DATA FURNISHED BY WAREHOUSEMEN. 23 Fresh milk: Minimum time of storage, in short storage of 1 or 2 weeks. Temperature of storage, 33° to 34° F. Buckwheat flour: Minimum time of storage, | month. Maximum time of storage, 6 months. Usual time of storage, 6 months. Temperature of storage, 32° F. Horse radish: Minimum time of storage, 1 month. Maximum time of storage, 6 months. Usual time of storage, 3 months. Temperature of storage, 33° to 35° F. Macaroni: Usual time of storage, from 1 to 4 months. Temperature of storage, 33° F. dry. Oleomargarine: Maximum time of storage, 6 months. Usual time of storage, 3 to 6 months. Temperature of storage, 0° F. Pickles: Usual time of storage, 3 to 6 months. Temperature of storage, 32° to 34° F. Watermelons: Maximum time of storage, 30 days. Usual time of storage, 10 days. Temperature of storage, 40° F. Pickled salmon: Minimum time of storage, 1 month. Maximum time of storage, 6 months. Temperature of storage, first 3 weeks, 28° to 30°; balance of time, 34° F. In some cases letters were written to accompany the blanks. A cold-storage company from the extreme Northwest writes as follows: * * * Judging from your letter you are in quest of data in relation to refrigerated products, not frozen meats, etc., such as we handle. Our business is that of wholesale and retail meat dealers. We store frozen animal and fish products and retail to the trade and to the consumer direct. We do not handle refrigerated meats at all; by that I mean meats that are held at a temperature but slightly above the freezing points for such products; all our stuff is hard-frozen, and is kept so until finally deliv- ered to the consumer. This is made necessary owing to transportation conditions which necessitate our laying in a sufficient stock to last through the winter months during the time of the open season in this country; 1. e., June, July, August, and September. We can not hold “cooled” meats that long, about seven months, and so we freeze everything ‘“‘hard.’’ That chilling meats improves them I firmly believe, but that freezing them adds to their quality I seriously question. During the summer months we furnish fresh-killed meats to the trade, but from October until the middle of June we handle nothing but the frozen products. We have had meats in storage as long as 14 to 16 months, at a temperature ranging from 14° F. in the summer, to 10° to 20° F. below zero during the winter, and they have been 24 PRELIMINARY COLD STORAGE STUDIES. furnished to the trade in apparently as good a condition as when they were first put in storage. We have no trouble in holding meats for eight to ten months in a hard- frozen condition, and the quality is eminently satisfactory. Very rarely do we have any complaints from the trade; once or twice during the season, perhaps. Given a stable temperature of about 15° F., with no more fluctuation than 2°, I believe meats can be held hard-frozen indefinitely, say several years, without further deterioration in quality other than that which transpires when same are first frozen. But in this country, where the outside temperature ranges from zero to as much as 60 below at times, it is impossible to keep an equable degree of cold, as the outside temperature is bound to have an effect on the cold-storage chambers, producing in them lowering of the temperature, and this excessive coldness has a tendency to take more or less of the life out of the meats and renders them slightly dryer than otherwise if kept for any length of time, say more than a year anda half. * * * A cold storage warehouseman of many years’ experience makes the following statement: * * * My present cold-storage occupation will not be of much use to you, as we do not handle any goods first-handed. I established one of the first cold-storage plants in the country and have had no end of experience, especially in the meat line. I have slaughtered at home, bought at Chicago both alive and dressed, and cut up all grades and qualities. Asa matter of trade cold storage is required to do this. After 48 hours it never improves the taste of any goods or their health-giving nutriment. I have eaten meat I have kept a year; while tender, it lost a large part of its taste. I do not consider it wholesome. I could fill 20 pages with experience, but it would not help you. There are so many different qualities in the same kinds that it would be hard to set a limit. Another storage company sends, in addition to the blank filled out, the following information: * * * We do not exactly understand whether the minimum and maximum time of storage is intended to show the customary times which these products remain in storage, or whether it is the intention to show the time certain goods might attain their best condition, and the time when they would be expected to begin to deteriorate. We presume, however, that it is the former, and have made our answers on that basis, assuming to a large extent that the withdrawals of these goods indicate the latter. Market conditions always enter into the matter also. Taking, for instance, fresh meats, we handle a great deal which is taken out almost as soon as thoroughly frozen, but the great bulk of it will remain for some time. We do not consider, however, that it can be carried satisfactorily over three months, as it will begin to show more or less dryness, etc. As to fish, it is not uncommon for us to carry them one year or longer, but we consider them in nearly all cases beginning to deteriorate after three or four months, unless they are redipped in water, thus forming a thin coat of ice over them which will protect them from deterioration almost indefinitely, so long as this process is renewed from time to time. Practically, however, this can not be done often on account of the cost, which — would become an important factor, and we merely refer to this as indicating what might be done. As to butter, where we show the minimum time of ten days you will understand that this refers to butter which is put in refrigerating rooms for the purpose of carrying it for a short time when the market may be overloaded or for some similar reason. For June creameries intended for use during the following winter the temperature is held from 5° to 10° above and also to 5° below zero. It is now assumed by the trade that the best results in carriage are obtained at the lower temperature. * * * INCEPTION OF THE WORK. 25 INCEPTION OF THE WORK. Among the most important of the foods so far examined on which the work is sufficiently advanced to warrant publication are chickens, quail, milk, and eggs. The conditions under which the eggs were pur- chased and stored are given under the appropriate caption on page 27. The preliminary study on milk, conducted by M. E. Pennington, has been reported in the Journal of Biological Chemistry, April, 1908, and will not be reprinted in this report, save for the conclusions drawn therefrom, which will be found in the summary on page 101. ~The chemical analysis of the eggs was made by F. C. Cook and the microscopic examination by B. J. Howard. The bacteriological ex- * amination of the eggs, quail, and chickens conducted at Washington was made by G. W. Stiles, and the study of market cold-storage chickens, covering chemical, bacteriological, and histological changes, was conducted at Philadelphia by M. E. Pennington. In the tests made at Washington a large number of chickens of as nearly as possible the same age and condition were secured in a perfectly fresh state, slaughtered, and prepared for storage by the removal of the feathers, and packed six in a box, as is the custom in placing samples in storage. The fowls were placed in cold storage at a temperature from 2° to 8° F.— that is, frozen solid—half being drawn and half left in the undrawn state. A sufficient number of these chickens were dressed, cooked, and subjected to the judgment of the jury in regard to the quality and character of the flesh. Another lot, representative in character, was selected for examination. A large number of quail in like manner were secured by special arrangement with the North Carolina game warden, so that the birds might be shipped directly to Washington on the date they were killed. In this way the exact length of time which had elapsed since their slaughter was known. ‘These birds were treated in the same manner as the chickens, having been tested and placed in cold storage, half drawn and half undrawn. At stated intervals, usually of about three months, samples of these storage products were withdrawn and sub- jected to the same examination as has just been indicated. Fresh chickens, as nearly of the same size and character as those in cold stor- age as could be secured, were always prepared with these samples. When these chickens were cooked and presented to the jury they were numbered or indicated in some way so that no member of the jury knew which lot was the fresh chickens, which the drawn chickens in storage, and which the undrawn chickens in storage. Each individual member of the jury was requested to determine for himself the char- acter, odor, color, and taste, to enter the same in a notebook, and to determine from the results of his notes and his experience which he considered to be the best for eating purposes of the samples presented to him. The members of the jury were not allowed to consult among 26 PRELIMINARY COLD STORAGE STUDIES. themselves until their final verdict had been rendered. The data thus noted were collected and summarized in order to determine whether or not there had been any consensus of opinion respecting the impor- tant points to be considered. A careful organoleptic examination of the chickens was also made before cooking, but in this case, inasmuch as the chickens were seen in the state in which they had been removed from storage, it was, of course, perfectly easy for each inspector to determine which was the fresh, which the drawn, and which the undrawn bird. Theseinspections consisted in:a careful study of the external appearance of the chicken, and also of the body cavity, in both the drawn and the undrawn chickens after the latter had been dressed for cooking. It is perfectly easy in such cases to distinguish between a fresh and a cold-storage chicken by the external and internal appearances. The odor, external and internal, was also noted. The recession of the eye of the cold- storage chicken, the changes of the color of the blood, the change to a certain extent of the color of the interior of the bird and its flesh, the development of blueness around the muscles of the legs, the ease with which the flesh is detached from the bone, the odor of the bird, the shriveled appearance of its skin—all are highly indicative of the changes which have taken place. It may be said that a careful inspec- tion of cold-storage fowls, whether drawn or undrawn, before cooking would do much to destroy any appetite which might otherwise have been manifested for these birds when cooked. The process of cooking, however, does very much to eliminate the differences between the fresh and the cold-storage fowls. To such an extent is this true that at the end of the first three months there were very great differences of opinion among the jury regarding the identity of the samples tested. It is generally considered by connoisseurs that a certain delay be- tween the killing and the consumption of chickens and game birds is necessary to develop the proper tenderness and flavor of the flesh. This is a problem, however, which can not be discussed very exten- sively at this point, since in the old-fashioned way the hanging of poul- try and birds for a certain length of time was considered necessary for the purpose of ‘‘ripening,”’ so called. They were not subjected, asa rule, to as low a temperature as that found in cold-storage warehouses. The common method, especially in Germany, was to hang theChristmas goose or turkey from a back window for ten days or two weeks previous to the serving of the meal, and thus it was subjected sometimes to temperatures below freezing and sometimes to those above. At the same time the birds thus exposed were hanging in the free air and were relieved of the danger of being left in the stagnant air of a storage warehouse. It is said that one of the best methods of sending the wild duck across the water in the winter is to nail them to the mast of a slow-going vessel. Their exposure in this way is said to place them COLD-STORAGE EGGS. OAT in the best possible condition for consumption on their arrival on the other side. It does not follow that the detention of birds in a closed cold-storage compartment below the freezing point for the same or greater periods of time will produce the same effect. I. COLD-STORAGE EGGS.“ PLAN OF WORK. The eggs for this investigation were secured from a reliable local dealer, who stated that all were laid on May 23,1906, and they were placed in storage on the following day. The wood of the crates was thoroughly seasoned and, because of absorption of odors by the eggs, they were made of white wood, gum, or other odorless wood. The strawboard supporting the eggs also affects the odor somewhat. All of the crates were new. The eggs were divided into 10 lots of one dozen each for storage and the weight of each dozen taken. One dozen was reserved for immediate analysis; the others were placed in a storage room having a temperature of 33° F. The weights per dozen on May 24, 1906, were as follows: Grams. Grams MICO NO De eo te a leila 6 ace GOOm Dozen: NOwieseoe2 seh oe: eek 2. 647 MnvemeNOn So) ick eee ee Bee Camm DoZemeNOnSespcs sees feos 2 . 647 ACTIN N Orch ae stew ees i CASE WozemeNon OMe i. Wier ee Sey un uy 612 ETN Os) Ses nae ears See ss te Sie ee CHO WOZENEINGO 2 Oe ese een ears Say 611 MATT OSE OR eee ge ee ees GAdee Dozer INO eal sce eae ne 623 At certain intervals, as indicated in the table, a dozen eges were removed for analysis. Four of these were examined chemically in a raw state and four were hard boiled; two were used in an attempt to determine volatile sulphur, but as only minute traces were found in either fresh or cold-storage eggs this estimation was excluded. One or two eggs were referred to the bacteriologist for examination, whose findings are given elsewhere in this report. , Samples were sent also to the microscopist. The following determinations were made on whites and yolks, separately, of both boiled and unboiled eggs,’ the weight of the material having been determined before and after cooking: Moisture, ash, ether extract, total sulphur, total phos- phorus, and lecithin phosphorus. The total nitrogen was deter- mined, and also the nitrogen in the form of coagulable proteids, vroteoses and peptones, and amido bodies on the boiled samples. The moisture, ash, ether extract, total sulphur, and total phos- phorus were determined by the methods of the Association of Official Agricultural Chemists.¢ The lecithin phosphorus was determined «Chemical analysis made by F. C. Cook. 6 The separation of the whites and yolks of the unboiled eggs was not satisfactory, and therefore the results were recalculated to the whole egg. See table, page 31. ¢U.S. Dept. Agr., Bureau of Chemistry, Bul. 107, Revised. 28 PRELIMINARY COLD STORAGE STUDIES. in the same sample in which the ether extract was estimated. The sample was extracted eight hours with absolute alcohol directly into the ether extract flask and the total phosphorus estimated in the ether-alcohol extract, which is computed as the lecithin phosphorus. The total nitrogen was determined by the official Kjeldahl-Gunning method @ and the amido bodies by the tannin-salt method.’ The proteose and peptone figures were determined by difference, the uncoagulable nitrogen minus the amido nitrogen giving the proteose and peptone nitrogen. The reaction which was determined in some of the samples was obtained by titrating a dilute solution of the whites and yolks with decinormal sodium hydrate or decinormal sul- phuric acid, using phenolphthalein as indicator. The coagulable pro- teid figures for the boiled samples were obtained by boiling 20 grams of the sample with 300 cc of water for three minutes, esti- mating the nitrogen in an aliquot of the filtrate, and deducting the figure thus obtained from the total nitrogen. The variations in weight referable to storage are given in the table. Loss of weight per dozen eggs placed in cold storage May 24, 1906. | Original) Final ea Time of storage. | weight. | weight. Loss in weight. 1906. Grams. | Grams. Grams. | Per cent. September(j:6-5) Months tases ese ea eee ee ee 623 |. 602 21 3 1907 | JANUBLY feel .0 EO ONGHS essa yars te ee ye ee eae eee eer 611 | 578 33 5.4 JUNMenl Al: GM OnTHS see Sec Saas ee ec Secs eee oe ee eens 612 565 47 ont October 61 6'6smonithss ee eee eee ee eee | 647 | 582 65 | 10.0 1908. | | January.14, 19:6) oniGhs ee eee eee ee een es | 647 582 65 10.0 } { In all of the analyses of cold-stored eggs, the samples were removed from the cold-storage rooms in the afternoon and immediately placed in the ice box at the Bureau of Chemistry, where they stood over- night, and the analysis was begun the following morning. The eggs, therefore, did not stand at room temperature for any length of time. What would have been the effect on the eggs of room temperature for two or three days, as is often the case in the household, these ex- periments do not show. PHYSICAL CHARACTERISTICS. Though the eggs had been kept at 1° F. above freezing, they showed a marked tendency to sweat when transferred to ordinary tempera- tures, the shells becoming very wet and also more brittle. After breaking the shell and keeping at room temperature for one day, the odor of the eggs in storage for 3.5 months was not unpleasant, aU. 8. Dept. Agr., Bureau of Chemistry, Bul. 107, Revised. 6 J. Amer. Chem. Soc., 1906, 28: 1485. COLD-STORAGE EGGS. 29 though it differed from the fresh. At the expiration of 7.5 months the difference was more marked, and after 12.6 months it had so increased that it was characteristic of cold-stored eggs, and can only be described as such. After 16.6 months a musty odor was noticed as soon as the egg was opened. The integrity of whites and yolks was preserved at the end of 3.5 months; at the end of 7.5 months considerable difficulty was expe- rienced in separating the whites and yolks of the unboiled eggs, though in the boiled eggs this was accomplished readily. When stor- age had been maintained for 19.6 months it was found impossible to separate the yolks and whites of the raw eggs, the latter invariably contaminating the former. The whites, which had been gradually losing their thick, gelatinous condition and becoming more and more _ watery, were now very thin and showed no tendency to adhere to the yolk or to cohere. The yolk membrane was so tender that when separated from the white it ruptured, though most carefully manipu- lated. When cooked, the whites were rather pink and the yolks were much darker in color than were those of the fresh eggs. The consist- ency of the boiled yolk varied from that of the fresh yolk when stor- age had been maintained for 16.6 months, the mealy character being much reduced. Neither did the unboiled yolks float on water, as they do when fresh. The physical variations from the normal are confirmed by the changes in weight of the eggs when cold stored as compared with the fresh. When fresh eggs are boiled, a loss in weight occurs, while storage eggs gain on boiling. Apparently the whites lose more water than do the yolks, and, consequently, gain more when boiled. The boiled yolks when fresh contain less than 50 per cent of water; when cold-stored this percentage is increased, the figure reaching 64 per cent in the last examination. CHEMICAL ANALYSIS. DISCUSSION OF DATA. The average ash content of all the samples of fresh eggs is slightly lower than that of the cold-storage eggs, and this is not entirely explained on the basis of the decreased moisture content of the cold- storage eggs, as may be seen from the table calculated to a water- free basis. The total sulphur and the total phosphoric acid in the boiled yolks do not show any progressive change for the eggs which have been in storage for a period of 19 months. After 34 months’ stor- age the percentage of total phosphorus as lecithin phosphorus in the boiled storage yolks is lower than in the boiled yolks of fresh eggs. It is probable, therefore, that the organic phosphorus compound of the egg yolk is being broken down into inorganic phosphorus. 30 PRELIMINARY COLD STORAGE STUDIES. The nitrogenous substances present in the boiled eggs examined show a change in the amount of coagulable proteid, which decreases during the storage of the eggs. The analytical data on the sepa- rated whites and yolks in a cooked state show a rise in the water | content of the yolks of the storage eggs and a decrease in this con- stituent in the case of the whites. The amido-nitrogen figures show a tendency to decrease in the case of the cold-storage eggs, but this effect is not uniform, and con- sequently no definite conclusion can be drawn as to a change in amount of amido nitrogen in cold-storage eggs. The proteose and peptone nitrogen content of cold-storage eggs shows a tendency to ‘increase. The reaction of the yolks, both fresh and those stored for 19.8 months, were slightly acid, while in both cases the whites gave an alkaline reaction. This acid reaction of the yolks of the storage eges was about one-half as strong as in the case of the fresh eggs. CONCLUSIONS. Eggs in storage for one year show a loss of weight equivalent to 10 per cent of the total weight, which loss is largely water from the whites. Eggs after storage for 16.5 months lose their power of cohesion and emit a characteristic musty odor a few hours after opening. A lowering of the amount of coagulable proteid of the boiled sample is indicated, as well as a change in reaction and a lower percentage of lecithin phosphorus in the storage eggs. An increase in the lower nitrogen bodies, proteoses and peptones, accom- panies the decrease of coagulable nitrogen in the boiled samples of storage eggs, while there is apparently a tendency for the amido bodies to decrease. Bul. 115, Bureau of Chemistry, U. S. Dept. of Agriculture. PLATE |. 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B : . : ‘opray |-ded puw| Stet | el | -ieqion, | - | -nsvog °0%q inydjns | ‘qovr3.xe ; : : ON sesoojJolg e101, 1e404, Tou1 Gt UusSW dINISTOW a[dures Jo uol4.d110seqy 218d retleg —sB Jueseid 889 JO UsSOIIIN “SISVE LHM ‘shba abp.0js-pjoo pun ysaif fo savy pajiog fo sishjnuy 49078—Bull. 115—08——3 34 PRELIMINARY COLD STORAGE STUDIES. BACTERIOLOGICAL EXAMINATION. TECHNIQUE. Ten fresh eggs and eight cold-storage eggs were examined for the number and the species of organisms present. The technique of this examination was as follows: The eggs were washed in a solution of bichlorid of mercury 1 to 1,000, or 5 per cent carbolic acid, for a few minutes, after which they were dried with sterile absorbent cotton and placed with the large end uppermost in a small beaker. The air space was then scorched with a gas flame for a few seconds. - An opening was made immediately into the cavity with sterile for- ceps, a sufficient amount of shell bemg removed without rupturing the membrane below. When this was accomplished the latter was broken with a hot platinum spatula and with a sterile pipette 0.5 cc © of the white of the egg was quickly removed and placed in the neces- sary petri plates for cultures. The remaining egg white was then decanted, leaving the unbroken yolk in the shell. With another sterile pipette the yolk sack was ruptured and suitable portions of its contents were removed for study. While such a procedure guards against contamination, the breaking up of the respective layers of the egg when out of the shell is difficult, and sometimes the inability to do so interferes seriously with the obtaining of quantitative results. With the eggs which have been in cold storage for consid- erable periods a separation of the whites and yolks is not possible. The first examination of cold-storage eggs, which was made at the expiration of the three months’ period, showed white and yolk still separate and the limiting membranes in a fair condition. At the end of six or eight months, however, there was an entire change in the character of the egg, the yolk and white having blended, so that it Was necessary to examine the total egg rather than its constituent layers. DISCUSSION OF RESULTS. The accompanying tables indicate the results of these examina-— tions. It will be observed that the fresh eggs had originally but few organisms, though in two cases, notably in the yolk, they were fairly numerous. The fact is also noteworthy that the most abun- dant growth occurred with the fresh eggs at body heat, and later at 15°-20°C. As far as the kinds of organisms are concerned, fungi, yeasts, and bacteria of various types were present. As one would expect, they were mostly air-borne organisms and of the types existing in the soil, cereals, water, etc. At the expiration of three months in cold storage, as will be seen in the following table, a very great increase in the number of organ- a Made by G. W. Stiles, jr., in cHarge of bacteriological chemistry, Washington, D. C. Bul. 115, Bureau of Chemistry, U. S. Dept. of Agriculture. Fi@. 2.—WHITE YOLK OF FRESH EGG (250). PLATE II. Y 4 % ¥] isms was noticed for both white and yolk. COLD-STORAGE EGGS. 30 The species isolated here are also of a great variety and fairly numerous, as the following Streptothrix sp.?, B. vulgatus, Sarcona lutea?, Alter- naria sp.?, Sarcina flava, Mic. ae Cladosporium sp.?, Mic. aerius, + 4 list indicates: _ B. mesentericus, Bact. ferrugineum, Sarcina subflava, and Mic. aureus. From this time on until the final examination—which was made resistant fungi. permit the drawing of elaborate conclusions. when the eggs had been in storage 1 year, 7 months, and 22 days— they were oie sterile or eeved by a few colonies of the more The number of eggs examined was too small to The observations are of interest, however, as indicating a line of future investigations. Bacteriological examination of fresh eggs. Number of Namnen of colonies per cubie | developing | developing on lactose | on lactose g 30° to 37°C. | 12° to 15°C. gelatin at wo SCoocoocorococoowrorcoe Mic. cinnabareus, Flugge. Sarcina lutea. Mic. cumulatus ? Bact. haematoides. ; Micrococcus, not classified. B. mesenter:cus. Fungus. 1 fungoid colony. 1 gray fungus. Actinomyces. a Gelatin plates incubated at 15° to 20° C. Bacteriological examination of cold-storage eggs. ret colonies eared ; per cubic i No. of | for ox- | Portion of | centimeter | centimeter sample. | smina- egg. tion. BER AG 5/24/ | White.... 1 pOGT. == -- ~~ - { 1906 | Yolk...... 190 : 5/24/ | White .... 0 <= { 1906) Yolk... .. 0 : 5/24/ | White 1 2069. ..-.-.- 1906 Yolk So 0 5/24/ NG Gravee il ee 1906 Yolk tec 240 5/24/ | White .... 0 a -- { 1906, | Yolk... 5 5/24/ | White 0 ek 1906 | Yolk. ..... 6 mmeeremi 1 Wry | White. 5 : 10/17/ | White 0 B.C. 552... { 107) | Yolk i 1/16; | White ....- 0 B.C. 669 4. { 1008) | Yolk. p ; 1/16/ | White 0 BC 670... { 1908 | Yolk...... 0 No. of sample. Duration of coid storage. LIS RSS Sie eee SPILT OS Sete Maye er als ee 2 eS SATU OSS, 2 eS tet ep aa eee LEAS SE eS ee eee GeOUMOSH Pets cote es Sees ke 1255) 26ers PSNI) Sisters fro ered pected ne ERRNO DE soe lek ee ERGO Se ee ae ee ae ey ee EN eS 34,0 SS = Sle, LUN e) Aa YO FS) eae eee en 2 OR / ae lyr. 7 mos. 22 days PERO ROOS cc oe sin oie 1 yr. 7 mos. 22 days Saccharomyces, B. Pammelii. Species of bacteria isolated. Muobacterium avium, Streptothrix farcinica, Portion of egg. Number of colonies per cubic centi- meter develop- ing at room | | | temperatures. | | eee HT aI Tea RONEN 6, 250 NG) ikea yee at ee Ee ee ee ee ea 3,370 NI VaIE Cem eens iin 2 ae, aoe 1, 280 WIEN aS eee ik, cae Ph oe ren Le 3, 260 A Ot ALC yee eee a eee nines 0 Motaepo tesa Soe eae ee oe 1 eR OG ACR oes. Suite as cay soe Soe eee 1 fungus. | MGtalles ene cw & ccen Lee esas cee White fungus at ice- box | temperature. NVALdUh 1 eS oes See ee Rie eer Atta 2 1 colony. INNO Kors Sa See eS ge itis See eee Slat 1 colony. | wee Ae eee ioe SRE ose ee AO ee 0 IONE 5. 2e eee eS on Polen a eae ere 0 ae ered ae 36 PRELIMINARY COLD STORAGE STUDIES. MICROSCOPICAL EXAMINATION.« FRESH EGGS. A hen’s egg consists of a shell, two shell membranes, the albumen or white of the egg, the yolk, and vitelline membrane. The shell ~ membranes consist of interwoven fibers (Pl. I). Those of the thicker ~ outer membrane are much the coarser (PI. I, fig. 1). At the broad end of the egg the shell membranes separate to form an air chamber, which increases in size as the moisture of the egg evaporates. — The albumen, or white of the egg, is fibrillar in structure and con- ~ sists of a dense portion next to the yolk and an outer, more fluid part. © The yolk is formed of alternate concentric layers of yellow and ~ white yolk. The yellow yolk forms the greater part and consists of ~ large spherical granular masses 20 to 109 in diameter (PI. II, fig. 1). These masses when hardened in situ are polyhedral, owing to equal pressure (PI. Til, fig. 1). The white yolk is found in a flask-shaped mass in the center of the yolk, and in thin concentric layers which — alternate with thicker layers of yellow yolk. The structure differs ~ from the yellow yolk in having spheres of several different kinds i which are smaller than those of the yellow yolk 40 to 80/in diameter — (Prieto: 2). The vitellme membrane surrounds the yolk and consists of -very fine fibrils. COLD-STORAGE EGGS. The first eggs examined in the cold-storage experiment were six fresh egos. These were normal when examined May 24, 1906, and it was stated that the eggs were laid the day before. The next exami- nation was made on a withdrawal of four eggs from storage, Sep- tember 7, 3.5 months later, and nothing abnormal was noted, the same being true of the withdrawal of January 7, 1907. Another examination was made of the June, 1907, withdrawal, representing a storage period of 12.6 months. At this time two eggs were examined and appeared in general to be normal eggs, but in the yolks were found small rosette crystals. It seemed that they were most fre- quently found in the outer portion of the yolk. In one egg they were of very rare occurrence. This was the first appearance of these forms and they figured later in the description of the samples of the - withdrawal of October, 1907, the last examination made. At this time the entire white of the egg looked watery; the part next to the yolk, although denser than the outer part, had deteriorated. The shell membrane appeared microscopically normal, as well as the. vitelline membrane. The yolk assumed a much flattened form after « By Burton J. Howard and E. A. Read, Microchemical Laboratory. Bul. 115, Bureau of Chemistry, U. S. Dept. of Agriculture. PLATE III. Fig. 1 —YELLOW YOLK OF FRESH EGG, HARDENED IN SITU (X250). Fia. 2.—ROSETTE CRYSTAL FOUND IN YOLK OF COLD STORAGE EGG (X250). QUAIL COLD-STORED UNDER KNOWN CONDITIONS. of removal from the shell and did not retain the spherical shape of a fresh egg but in all cases was distinct from the albumen. In addition to the normal yolk elements, rosette crystals were found in the yolk. These measured from 18 to 63/ in diameter (PI. III, fig. 2). The exact nature of these crystals has not yet been determined, but the following points were established: (1) The crystals are soluble in warm glacial acetic acid but are insoluble in ether, alcohol, or cold mineral acids; (2) the crystals melt and recrystallize when stip acids are heated. ' SUPPLEMENTARY WORK ON ROSETTE CRYSTALS. Two eggs were examined about February 15, 1908, when they had been in storage approximately 20 months. In general appearance the eges were the same as those withdrawn in October, 1907. The rosette crystals were larger and more numerous than those previously found, some of the crystals measuring 109/. The study to determine the chemical nature of the crystals is still being carried on, but the difficulties of the problem make the progress very slow. iene have been made to separate the crystals for examination, but thus far with only partial success. > 7" DEGETO DOL SAT Ge | ee 55 2 see ae aici as IOpO pusv o94S¥], | -oJul ‘ssotoqseqy fAIq |- 77> > -1vedde 4seq—UM BIC “1OOMS SSO] “poo “100d AIA *poos YONuUL 998B4 {T “ON “g0uBIved SO Ones OO Cee LONE ieniesLOAGG [ae O Ulam |) UL ae AeT (ale | me aia at lea IOpO ‘!3U019S 99SB], | SB ISBMS SB IOU IOPO | 7 7 e -d¥ 4s1IOM—UMPIPUL) *poos 40U 10po “AIP -gouvived “Ove, CAMO, || Bohewien — jowit) jGovaysy_ IOP SeO SSS Seabee sanconGe “poo3 1opo puv oysvy, | QooMSIOPO puBoysBy, [~~~ -de 4SIOM—UMBIC “(ADVUOLS NI SHLNOW 8) 9061 “Sl UAAWAAON “Sp1Iq "109MS DUB ‘LON ysely uvg} elip ‘aig ‘yivp ‘Arp ueyqi Auies os10Ul 489UI YIVpP !pooy q10q ‘7 ‘ON ueyy 19490MS JBMOUIOg “ron ‘Va0MS (HDVUOLS NI SHLNOW &£) 9061 ‘¢ ANOS p89} WNC) {BOUL ONIUM ST “ON "1OABT "TON OXI noqe sve o[qeyvyed OS JON | Sse] ‘TON O41] ody: ‘poos LOpO puB o4SBY, |---- UMBIPUY) “AUB “IOABVY Poos elouL ‘{UO0T1eIOTI04 “WOILBIOLIIJep OU ‘uly Ale | -op ou {Arp ynq ‘poosk) | :poos LOpo pu oysBy, |--- >> UMBIC. “aL WN iclesdlamva SANK AMA “IE “AO 40 PRELIMINARY COLD STORAGE STUDIES. BACTERIOLOGICAL EXAMINATION. The bacteriological examination of quail was conducted along the same lines as was the examination of cold-storage and fresh chickens. As before stated, they were killed especially for this work. Half of the number was drawn, the other half left undrawn, and all placed in the freezer as promptly as possible. The feathers were not re- moved. Thawing was ordinarily allowed to proceed at 10° to 12° C., after which the examination was conducted as promptly as possible. Because of the difficulty in obtaining fresh birds at all seasons of the year, it was sometimes necessary to examine those in storage without having a fresh bird at the same time for comparison. When- ever possible, however, fresh birds were obtained, and in one case a bird which had been 3 weeks in storage was used as a basis of com- parison. Even so short a period, however, caused minor irregularities, hence the findings for this quail can not be accepted as absolute. In all, 4 fresh quail have been examined, 3 being from the same lot. No. 4272 was examined as the comparate of the birds in storage for 11.5 months. The other fresh quail were tested simultaneously with the birds in storage for 13 months. No. 4272 showed a number of bloody spots, which indicated the site of the shot wounds. It had also a broken leg and a broken wing. The body cavity showed that the shot had penetrated and had torn the viscera as well as broken the intestinal walls, which permitted the diffusion of the contents throughout the body cavity. Such a condition would of course infect any organ which had been damaged by shot. Those which were not so damaged were sterile. The species isolated, and their location, are listed in the table under sample No. 4272. Nos. 752 and 754 were in good condition except for shot wounds which had torn the viscera of No. 752. No. 753 was badly torn, both externally and internally, and was dark in color for a fresh bird. There was no marked odor or discoloration, yet decomposition had apparently begun. - The first examination was made when the quail had been in stor- age for three months. The external appearance of both the drawn and the undrawn quail was good. The body, however, was altered in shape from pressure while in the case. The undrawn was the better looking of the two. Both birds had a peculiar odor, not very strong but suggestive of rancid fat, which was more pronounced in the undrawn than in the drawn. Several small areas on the breast muscle showed evidences of desiccation, and in the drawn quail this muscle showed shot wounds, which were surrounded by a bloody exudate. The fibers of the muscle were brittle. a4 Examination conducted by G. W. Stiles. QUAIL COLD-STORED UNDER KNOWN CONDITIONS. 41 The viscera of the undrawn bird were in fairly good condition. A small amount of a dark extravasated bloody effusion was found in the abdominal portion of the body. There were no gas bubbles and no discoloration except where the liver came in contact with the parietal walls. | The body cavity of the drawn bird contained portions of the liver and kidneys. These were slightly softened and dark in color. The blood which was in the body cavity was quite dry. No particular discoloration of the tissues was noted and the bones appeared normal. As will be observed in the table under No. 4073, this quail had an infected brain, and also gave a bacterial growth from the breast muscle in the region of the shot wound. This muscle, however, near the crop where the skin was unbroken, was sterile. The only organ- ism isolated in pure culture was Micrococcus tenacatis. In the undrawn quail bacteria were found in the leg muscle. The leg, however, had been broken; hence it is possible that the infection was local. The organisms isolated are given in the table. The second examination of quail preserved by cold was made after a storage period of 8.5 months. In both birds the odor was suggestive of rancid fat. In the undrawn bird, however, there was a distinct fecal odor in addition. The region of the vent in both birds was darkened in color, with rather a greenish tinge in the undrawn. The muscles of both birds cut very readily. The abdominal cavity of the undrawn bird contained about 10 cc of a bloody serous fluid. The kidneys were dark and congested; the intestines of a dull gray color not at all attractive in appearance. There was also a noticeable odor of putrefaction when the abdominal cavity was first opened. The membranous lining of this cavity had a slimy appearance and a greenish-yellow tinge. In the drawn bird the thoracic portion of the cavity was in very good condition, but the posterior region was abnormal both in appearance and in odor. The lining membrane, particularly in the region of the vent, was infiltrated with a slimy, bloody fluid. The crops in both birds contained the seeds composing the food of the quail, and a few gas bubbles were found entangled within the loose connective tissues surrounding it. The muscle was apparently in good condition, the only discoloration being where the shot had penetrated. The only organisms isolated from these birds were from the intestines, cultures having been made from various regions as indicated in the table, where the quail are numbered 4270 and 4271, respectively. The next examination was made when the quail had been in stor- age for 11.5 months. The general external appearance of both the drawn and the undrawn birds was bad. The body was distorted in form and broken bones were noticed in both cases. The muscles 42 PRELIMINARY COLD STORAGE STUDIES. were shriveled and the region of the vent was distinctly sunken, varying in color from green to a brown-black, the discoloration extend- ing well into the tissues of the inner side of the leg. The skin was parchment-like. All of these conditions were more pronounced in the undrawn than in the drawn quail. The odor was putrefactive, rancid, and fecal for the undrawn, and distinctly rancid for the drawn bird. The muscles of the drawn bird were in better condition than were those of the undrawn. The texture of the latter was flabby and without elasticity. In the undrawn bird the abdominal cavity showed the presence of mites. The heart was in fair condition. All the other organs were in exceedingly bad condition, the liver, for instance, being soft, light green or yellow, and much degenerated. The kidneys were dry and friable, varying from gray to yellow-brown. The intestines were green, the folds being matted together in masses without any distinctive outlines of demarcation, and the walls so thin that they broke even when lightly touched. This condition grew worse toward the vent. In the drawn bird the body cavity contained blood which was nearly black, the walls of the cavity being greenish-yellow near the vent. The lungs and kidneys were in situ, but so soft and darkened in color that their structure was almost lost. Cultures from these birds were made and the results are given in the table under No. 4431 and No. 4432. All the cultures made from the drawn bird were sterile. From the undrawn bird growth was obtained in a number of instances, and the species isolated were varied both in character and in location. The bird used for comparison (No. 4433) had been kept in stor- age for three weeks, this period being necessary because of the closing of the season for the killing of quail. The bird was in a very good state of preservation. However, the region of the vent showed a slight greenish-yellow tinge. There was a shot wound in the right leg, which still showed the shot embedded in the tissue, and the inner thigh muscles were discolored and hemorrhagic, this condition extending well toward the vent. The lungs were somewhat con- gested and the kidneys slightly darker than those of the fresh birds before examined. All the cultures made from this bird proved to be sterile. | 3 : After a storage period of 13 months, the two undrawn birds—Nos. 755 and 756—had discolored skins, especially in the region of the vent. The muscles were also discolored and dry. The viscera were in bad condition, dark in color, the intestine leaden, the tissues degenerated even to macroscopic observation, and a bloody, slimy fluid present in the body cavity. The odor was fecal. QUAIL COLD-STORED UNDER KNOWN CONDITIONS. AD The drawn birds of this lot—Nos. 757 and 758—were in a bad state of preservation externally and internally. The breast muscles were soft and degenerated, stained red where in contact with the bone, and the bone marrow was absorbed. The portions of the vis- cera left in situ were dark, bloody, gaseous, and slimy. It will be observed from even the small number of birds exam- ined, and here listed, that the problem becomes even more compli-. cated than in the case of the chickens, because the method of killing is almost certain to introduce bacteria which may be carried into the deepest tissues and, as in the case here noted, result in the rupturimg of the intestine and the pollution of the entire body cavity. The prompt dressing and cooking of birds so injured would render this factor unimportant. If, on the other hand, the birds are exposed for sale for a number of days before being disposed of, or being un- sold are, at the expiration of such a time, transferred to cold storage, the condition may be menacing. It would seem, in the case of quail even more than in the case of chickens, most desirable that a very prompt transference to the storage warehouse be made, or that the birds be dressed and sterilized by heat as soon as possible after killing. Bacteriological examination of fresh and cold-storage quail. FRESH QUAIL. Cultures. UNS) Oe) a No. | Description. storage. an Re- | Species isolated. sult. | Months. Apa Gawain sierra oe Heart bloods ae. eee Strep. pyogenes?, Aspergillus glaucus Staph. epidermis? Breast muscle near crop. - | Peritoneal fluid ........-- WG behenl. See eee WAN OT2 es earns Ae ons LiCSKG NYE 72ers Gs | Breast muscle near crop- - Strep. pyogenes, Mucor racemosus. 7524 Bact. salmoneum? 0 0 + + 0 ae + 0 0 | Weems Cle peer ae eee = 0 | IBONeMMaATTOWe =e oe see 0 TESTA 1 Uprave bite hy,0 eye | eee Hieamtablood a2. eee oe + Culture 737.0 + a + + + + 0 0 0 0 [KGL ine Wires Mee eae eee en Breast muscle near crop. . Culture 737.6 B. subtilis group. epumilscles ae shee 4) Una rawmes 2! oo. os Taare lokeyore! ee Culture 737.6 M. pyogenes albus. TIVE Dee ses fare hn ee ees = Kidney. . ae M. pyogenes albus. Breast muscle near crop.- ; RC ovIMUISClE SEA eae. oe eet BonenmMa Trower eee esse. = Renitoneal timideys soa se. re) "oO = Ler 4 ct sel = o) o) jon 4433 Undrawn....| Breast muscle near crop- - | Peritoneal fluid......-... 4073 | Drawn.....- 8) RS lSHH IS Se = eS ae ae | | Ricoriploods a wales | Breast muscle near crop. | | Breast muscle near + | wounds. M. tenacatis. cotocooceo a From same lot of quail, results comparable. 6 All cultures numbered refer to bacteria not classified. 44. PRELIMINARY COLD STORAGE STUDIES. Bacteriological examination of fresh and cold-storage quail—Continued. STORAGE QUAIL—Continued. | Cultures. No. | Description. eee evenness | Re- Species isolated. Ee sult | Months. 4074 | Undrawn 3 ABUT y Neen ees ee heey See 0 |- (same lot IEF OOO. scancasouc sue Ho X0)s as 4073). Breast muscle near crop.-| 0 | kecamusclesee sere ee eae + | B. cloacz, Spir. serpens. 427 n Dra wiles. = Reon (iol BIL eH OO ep ee ee te ee eee 0 TUM Ss hen seer eee 0 Breast muscle near crop 0 | IBOneeMaTLOWe + ss5 eee ae 0 2VATD | WrraiGliPeinaigals oe solos cl leimyine S352 ota e oko desde 0 (same lot EeGant blood meres see ates 0 as 4271). A VOTE apne cee ee EE 0) HRIGNCY Sete Sse ee (Oy ==) PGES TIMER aA tern eo + | B. circulans?, M. aurantiacus. Breast muscle near crop..| 0 | IBONGMMaLTOWse see ere eee 0 Peritoneal fluid: --.--2:2- 0 44319) maw 11 Soul Mele arta lOOd esa eae 0 Breast muscle near crop..| 0 | Breast muscle near 0 | wounds. | 4432) Und raw n!|5--2d0- 3) Heart) blood!s==22-. 2). -.- | + | Strep. erysipelatos. | (same lot DIVER eso Bes es et | + | M. tenacatis? | as 4431). ung sees Wet eras pee + B. enteritidis. Tanites tines ess ene oe + | M. tenacatis’, B. capillaceus. Rericardiaiiuida === ee + | Strep. erysipelatos. Breast muscle near crop..| 0 | Peritoneal huids. a2 sess Ors (ome Drawae 2.22 | 13 Breast muscle near crop.-| 0 | | esau SCle eee ee se 0 | | Bonemarrows 2.42 ess. Oia 758 | Drawn(same |....do..| Breast muscle nearecrop.-| 0 | lot as 757). Hegamiuscleseees sear OR IBOnNeEIMALTOWRes espe ee eee O-) (554 sO aawen|see- COee| Sr eantiblOOdm eae smear + | Feeble growth, failed to develop when (same -lot | IDR Cieaeiee re ao aes ee ae ace One) transplanted. as 757). | Intestine sehen See Om | Breast muscle near crop.-| 0O | | ibecsmuscleses= nt ae On} | BOneHNaLLOWAe eases ete 0 (567 | ain diarw ne 2se4d G24) sHeanrtibloodeet sess. saesee 0 (same lot | I Diy {e1ee eee ate Tee 0 | as 757). UGG AGN Ga ee aera ae + | Feeble growth, failed to develop when lin'teS Cine aa a ee 0 transplanted. Breast muscle near crop.-.| 0 hes muscle sees eee 0 One MaArtOWwe ase see 0 III. CHICKENS COLD-STORED UNDER KNOWN CONDITIONS. ORGANOLEPTIC TESTS. Tests were made on chickens placed in storage on July 1, 1906, and withdrawn at intervals of approximately three months for a period of almost nineteen months. At each test a drawn and an un- drawn chicken were taken out of cold storage and compared with a fresh chicken both before and after cooking, the jurors having no information as to the history of the birds and giving their opinions -independently. The chickens were cooked by the same person throughout the experiment and prepared in the same manner. ~On October 19, 1906, after three and a half months of storage, the cooked chickens showed practically no deterioration and the jurors found it a little difficult to distinguish them from the fresh fowl. CHICKENS COLD-STORED UNDER KNOWN CONDITIONS. 45 At the second test, however, on January 16, 1907, three months later, the fresh bird was easily distinguished by all of the jurors. This was also true in the other three tests, the difference becoming more pronounced at each trial, and only one mistake was made during the entire observation in the distinction between the fresh and the cold-storage fowls. About 70 per cent of the votes cast at the second test declared the undrawn fowl to be inferior because of its darker color and its stronger odor and taste. After almost a year of cold storage, June 11, 1907, the third test was made. The undrawn cold storage fowl was correctly designated by all except one of the jurors, and was described as being of dark color with a bad odor and strong taste. Four months later, in the test of October 22, 1907, when the birds had been in storage for 15.5 months, the distinction between the drawn and undrawn cold-storage birds was less pronounced, but there was no question as to which of the three birds was fresh. About 70 per cent of the votes again indicated correctly the undrawn birds, which were described as being dark and dry, the bone at the joints darkened, the white meat tasteless, and the dark meat strong in the majority of cases. When the last test was made, on January 23, 1908, the birds had been in cold storage for 18.5 months. honck the difference between the fresh and the storage birds was obvious, the jurors found it even more difficult than in the preceding test to discern the difference between the drawn and the undrawn storage chicken, the vote being evenly divided; both of the storage fowls were by this time somewhat dry and dark, and tasteless rather than strong. From the evidence of the jurors who examined the cooked birds, the following conclusions are drawn: The undrawn fow! is distin- guished at first with comparative ease, being inferior in appearance and usually stronger in odor and taste. When they have been — stored for 18 months, however, it is difficult to discern a difference in the two, as both are dry and tasteless. The jurors are practically never in doubt as to the superiority of the fresh bird, and this becomes more marked as the experiment progresses, though a short period of storage (3 to 6 months) seems to cause no deterioration. Besides the table tests, the fowls were subjected to a macroscopic examination before being cooked. When the chickens had been in storage for 6.5 months, the verdict was that both externally and internally the undrawn bird was better in odor and appearance than the drawn cold-storage bird. On June 11, 1907, the second examination was made after a storage period of nearly a year, and it was found that the internal appear- ance of the drawn fowl was still good, no serious decay having appar- ently occurred. The internal examination of the undrawn chicken 46 PRELIMINARY COLD STORAGE STUDIES. showed signs of degeneration of the organs, but the odor was not bad, though a little stale. Both fowls were in fairly good condition. After 15.5 months of cold storage, on October 22, 1907, the undrawn bird again presented a heres appearance een than the drawn one, both internally and externally. The flesh of the two fowls was somewhat discolored, but. the odor was not bad. The liver, the lungs, and the muscle tissue were all in better condition 3 in the undrawn chicken. On January 23, 1908, when the chickens had been in cold stoiage for 18.5 months, they were again examined. There was practically — no disagreement as to the inferiority of the undrawn bird in this test, both the internal and external appearance, and also the odor, - showing clear evidences of degeneration. The conclusion drawn from the examination of the uncooked chicken is that after 12 or 15. months of cold storage the undrawn chicken appears better than the drawn, while after 18 months in storage the undrawn chicken is in decidedly the worse condition of the two in every respect. Detailed descriptions of the appearance of the chickens at the last two examinations are appended and the results obtained in the organoleptic tests are comparable with the bacteriological findings of corresponding dates, such birds having been from the same lot. Macroscopical examination of fresh and cold-storage chickens, October 22, 1907, after 15 months of cold storage. IRR Op Fresh (black chicken).—Color of skin “white; tissues normal, soft and elastic with normal odor; breast muscles soft, yielding, and elastic, nearly colorless; thigh muscles none the various muscles being easily differentiated by color, muscle sheath, and elasticity. Liver firm, elastic, and of normal color; gall bladder normal; intestines firm, resistant, contour well marked, normal color, small blood vessels noticeable. Drawn.—Yellow color; odor not bad (too cold to test); tissues dry, firm, dehydrated, and adherent in places, though not to such an extent as the undrawn bird; breast muscles dry, moderately firm, striations not so marked, lighter pink color and better appearance than undrawn chicken; thigh muscles hard, dead appear- ance; colors of various muscles not differentiated. Liver and lungs hardly recog- nized from their gross appearance; heart not a bad color; gizzard fairly good condition. Undrawn (Barred Plymouth Rock chicken).—External color yellow; Sin not bad (bird too cold to test); tissue dry, firm, dehydrated and adherent in places; breast muscles moderately dry, firm, striations rather marked, dull pink color nearly red, not natural; thigh muscles hard and rather dry near external surface, dark red in color; muscles not clearly differentiated in color and muscle sheaths not well marked. Liver soft, faded and mottled color, with tissue neither natural nor elastic; gall bladder partly empty and pale green; gizzard a little darker than hota: heart not a marked blood color; intestines soft, mushy, and pale, all traces of blood vessels gone; fat rather yellow. CHICKENS COLD-STORED UNDER KNOWN CONDITIONS. 4 “I 5 OD oe Fresh.—Eyes full, comb and gills pale; neck soft and well rounded; feather papillee prominent. Region of crop inconspicuous, but not depressed. Body clear pale pinkish yellow color, with bluish tones over legs and hips; skin firmly but pliably attached to tissues beneath. Muscles of breast almost colorless, opalescent grayish pink; texture very elastic. Muscles on inside of thigh pink and soft; fat pale yellow. Dark, blood-red liver, well rounded intestines, blood supply distinct; gizzard bright. Drawn.—Head gone; end of neck powdery, dry; skin like parchment, adherent to neck, loose over breast, easily torn away from muscles, much too yellow; tissues below dried out. Muscles of breast too pink and dried, fibers very prominent, elas- ticity largely gone. Inside of thigh too deep in color, purplish and brownish tints appearing; fat too yellow. Gizzard, liver, and lungs loose in body cavity, hardly recognizable. Undrawn.—Eye depressed, comb and gills dried; neck shrunken, skin adherent. Skin better than drawn, but not normal. Breast muscles too pink, very fibrous, not elastic, not so dry as drawn and generally in better condition. Tissue of thigh better than drawn, but tending toward some coloring. Liver lighter in color and not resistant to pressure, mottled; gizzard normal; intestines flattened, veinings indistinct; color not bad. Gall bladder has discolored tissues around it. Fat slightly discolored. BCC. Fresh.—Skin light in color and tight; odor and appearance good; interior good norma color, with organs fresh; odor as in all fresh chickens. Drawn.—Skin rather dry and hard; odor and appearance good. Internal organs discolored and shrunk, but not of bad odor; muscle fibers dried and striations very distinct; odor all right. Undrawn.—Skin dry and hard; odor and appearance good; appearance of interior better than in drawn fowl; the organs fairly normal, only slightly discolored. Muscle tissues look better than in drawn chicken. Macroscopical exanunation of fresh and cold-storage chickens, January 23, 1908, after 18 months’ cold storage. Te WW Fresh.—Y ellow and pink flesh; purple and red legs; plump; pleasant odor; eye full; interior flesh and intestines red; odor fresh; liver brownish red; lung tissue yellow. Drawn.—Faded color; flint legs; shriveled skin; bad odor; head gone; interior flesh appears more nearly normal than that of undrawn; odor also better. Undrawn.—Faded color, pale, and shriveled; bad odor; eye sunken and almost invisible; red color of intestines gone; liver bright yellow, very bad odor; interior flesh faded and bad smelling; many evidences of degeneration. Lhe Gan UA Fresh.—Color normal pinkish; odor as of fresh meat; tissues soft, friable, and moist; eyes normal. Drawn.—Tissues dry, hard; color darker than normal; skin drawn and tightly adherent; odor better than undrawn; head off; interior tissues tough with no resilience; odor and color better than undrawn. 48 PRELIMINARY COLD STORAGE STUDIES. Undrawn.—Muscles dry and hard with skin tightly drawn and adherent: color dead and dark; odor stale; eyes sunken; interior tissues pale, hard, and tough, with no resilience, bad fecal odor; lungs mottled white; liver flabby; gall bladder half empty; gizzard smaller than normal; intestines soft, mushy, with no blood in the blood vessels and through omentum; blood vessels in walls have disap- peared. - He CNC: 5 Fresh.—Rather small chicken, 14 pounds; color and odor normal; eye full and round; interior color and odor normal; liver brown-red. Drawn.—Skin and muscles rather bruised; skin dry; not plump; odor good; interior color pale, but more normal than undrawn, and odor good, though not strictly fresh. -Undrawn.—Skin dry and bruised; shape lost owing to packing; eye has disappeared; odor good; interior color very pale; meat has lost its characteristic appearance; hemoglobin reduced. 49 CHICKENS COLD-STORED UNDER KNOWN CONDITIONS. *AIp puv ‘ysnoy ‘Su0I9S QeoUL OqTGM ‘9019 oy) JO 4ysesu0119 “ysnoy pue osi1vod J19yQeVI ‘T ‘ON S® poos sv jou ‘gouvivedde yusT[I0xe qysouL puBv YSoIy IOPO *[MOJ 058 -101Ss uMBip ATqeqord ‘QooMS puB Jepuey * ‘S]JouIs Jt UBYY 10430q soysey ‘fairey oouviveddy “Su014s AIOA ‘ROTI yiep jo Ayeroedse ‘91Svi ‘su019S J10po f10]OO UI [[Np ‘pepeaalys “IIBJ IOARY * 1 “ON Uv Jepue} VIOUL ‘9ATY -e30u 10poO ‘] ‘ON SB poos os jou vouvIroddy “Ysno} 19y4e1 f11Ivy IOARG ‘4o0MsS AIOA IOPO ‘SsByjo 4ST 'Z ‘ON URYY 10930q qnq ‘poos jou o4sv4 ‘peq Apysys z10po ‘Z “ON SB YSoly SB YON “ysoly 10U ‘99sB4 -199J8 pvq JVOUL 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ONG lsue eae Ysolj—]J “ON D'D MM “HH “LO 1D ICs Ul “Ua LW “AOS “(SHLNOW {S1) 2061 ‘2 UAAOLOO ‘ponutyuo;) "SYJUOUL dary) fO SPDALaZUL 1D Pajsa) pun payooo ‘suayoyo abp10js8-pjo09 fo WoYYpuUod pun ‘OPO ‘azSDT, ab CHICKENS COLD-STORED UNDER KNOWN CONDITIONS. 51 BACTERIOLOGICAL EXAMINATION.“ The bacteriological study of the fresh and cold-storage fowls has been concerned with the presence of bacteria in the tissues, and their species. The birds were sent to the bacteriological laboratory from a lot obtained for general work and, previous to their reception at least, were in nowise differently treated from the others. When received at the laboratory they were examined immediately, or, if this was impossible, they were placed on ice for the short period of waiting. The portions of muscle to be examined were obtained by searing the surface to insure sterility; then small pieces were removed with a stiff platinum hook such as is commonly used for this purpose. The fragments were transferred to suitable media and incubated at body heat, and in many cases also at from 15° to 20° C. The media used in routine consisted of plain agar, nutrient bouillon, plain gelatin, elycerin agar, and, for the detection of gas-forming organisms, a 2 per cent dextrose broth. Growth from the fresh chickens occurred with a fair degree of promptness. In the cold-storage chickens, on the other hand, six days or more were required at 20° C. A close examination was made of the physical condition of the birds, and brief descriptions of them are here given. The results obtained with the fresh chickens are tabulated in the table. These fowls were allin good physical condition. They were drawn, and had been picked. The muscles were elastic, yet tender. The birds were killed by puncturing the spinal cord by way of the mouth. Hence there was a wound in the head, and, owing to the manner of killing, it was of course more or less infected with organisms from the knife. Here and there there were slight abrasions of the skin and sometimes the wing bones were broken, such accidents being likely to happen in the killing and picking of the fowls. Internally they were all in good condition. Parasitism was not noticed in any case. The odor of the flesh was pleasant, lacking rancidity. The tissues in the region of the gall bladder had ee a greenish tinge. Hiro of the six fresh erideone sumed were sterile, cultures being made from heart blood, liver, spleen, bone marrow, and muscle tissue. All of the others showed one or more organs infected. The variety of organisms isolated, however, was not large. In this connection it should be noted that chicken No. B. C. 559, received shortly after killing, was kept in a sterile metal container on ice overnight. It was, therefore, somewhat more than twenty-four hours old when the examination was made. No. B. C. 694 was eRe on ice under the same conditions for a few hours only. The first examination of cold-storage fowls was aoe when they a coerced he é W. Stiles, jr. 52 PRELIMINARY COLD STORAGE STUDIES. had been preserved under the conditions previously noted for 119 days. An undrawn and a drawn specimen were examined simul- taneously, and comparisons not only with a fresh fowl but with one another were made. From the undrawn fowl only the feathers were removed. From the drawn, the head, feet, legs, crop, and intestines were removed. Originally the lungs, heart, liver, spleen, testicles, and kidneys were left in position, and the vizzard was emptied on returned to the body cavity. Externally both of these chickens showed some broken skin. Both had an odor, but that of the undrawn fowl was faint and not unpleas- ant, though rather different from that of the fresh bird, but distinctly better than that of the drawn fowl, where indications of beginning putrefaction were plainly evident—a condition observed for the undrawn around the head only. That portion, however, was dis- tinctly foul. In the undrawn chicken there was no apparent change in the color, while in the drawn there was a tinge of greenish yellow in fat and subcutaneous muscle. The viscera of the undrawn fowl were in very good condition, and the fecal odor noticed when the incision was first made soon dis- appeared. There was some bloody effusion present in the lymph spaces of the abdominal cavity. This was always noted for the drawn bird. The liver was pale in color, but of good texture. It was some- what green where in contact with the gall-bladder ducts. The tes- ticles and mesenteric fat were somewhat more yellow than in the fresh chicken. This also applied to the chicken from which the viscera had been removed. Cultures were made both from the organs and | from the muscles. The infected organs and also the list of species, and their sources, in the chicken will be found in the accompanying table under speci- mens Nos. 4251 and 4252, respectively. | The second examination of cold-storage chickens was made when storage had continued for 215 days. As in the former examination, there was a comparison of the drawn and the undrawn cold-storage ‘chickens with one another and with a fresh chicken. The undrawn chicken had a broken right leg, which was dry and darkin color. The | drawn chicken had a open een wing, which was: also dark and bloody, with an odor of parericior: The ee chicken had a strongly fecal odor in the region of the vent, which was less marked toward the anterior end of the body. The head, however, had a strong odor. In the drawn chicken the | odor was mildly rancid, somewhat butyric in character, and but | slightly putrefactive. The tissues of the undrawn chicken were soft, watery, and rather | slimy, both to appearance and to touch. The same condition was | CHICKENS COLD-STORED UNDER KNOWN CONDITIONS. 53 “noticed for the drawn chicken, which showed in darkened areas and | ce hE. | streaks here and there. . No difference was noted between the drawn and the undrawn in the texture of the muscle. In the undrawn fowl the spleen and kidneys appeared practically normal. The heart muscle was rather soft and the liver pale yellowish ‘in color and easily disintegrated. In the drawn fowl the lungs were ‘dark, bloody, and very unsightly. The heart, liver, and gizzard had been dressed and returned to the abdominal cavity. They were rather dry in appearance and somewhat darkened in color. The color of the hip bones and ends of the joints in this drawn chicken were much reddened and the subcutaneous fat, particularly near the ' vent, had taken on a greenish tinge. On the whole, the general phys- ical condition of the drawn bird did not compare asfavorably with the fresh chicken as did the undrawn. The bacteriological findings for these two chickens are given in the table as Nos. B. C. 4295 and B. C. 4296. The next examination of the drawn and undrawn chickens was’ made after a storage period of 384 days. These chickens were thawed jn an ice box protected by a sterile tin can closed by sterilized absorb- ent cotton. It was noticed that. the normal shapes of the chickens were decidedly altered by pressure in the packing box. The odor of the undrawn chicken was slightly rancid and some- what fecal. The colorcompared favorably with that of the fresh fowl, there were, however, discolored areas over the back and the sacrum and a few blebs of gas in the subcutaneous cellular tissue. An examination of the body cavity showed soft watery viscera mixed with blood and much degenerated. The liver was pale and soft. The other organs were rather darker in color than the normal. The intestines were grayish yellow, degenerated, and very distinctly different from those of the fresh bird. Except for the condition of the lungs, the body cavity of the drawn chicken was in better condi- tion than that of the undrawn. The heads of the long bones were dark red. As was to be expected from the macroscopic appearance of these birds, the bacteriological findings were fairly rich. They are given in detail in the table under Nos. B. C. 14 and B. C. 15. Chickens Nos. B. C. 560 and B. C. 561 were im storage for 517 days, There had been, since the first examination, an increase in the num- ber of infected organs, and, keeping pace with this condition, the number of species isolated increases. With such a bacteriological condition, there coexisted the following macroscopic appearance: The skin, in both the drawn and undrawn chickens, was somewhat discolored and becoming dry, especially on the back, tips of wings, and such prominent portions. In general, the tissues were plump, though the breast of the drawn chicken was drying out. The body 54 PRELIMINARY COLD STORAGE STUDIES. - cavity of the undrawn chicken showed a pale, jaundiced liver; mot- q tled grayish-yellow lungs; darkened kidneys and degenerated intes- — tines. In the drawn chicken the lungs, heart, and kidneys remained in situ. They were all of a muddy, dark red appearance and there was a thin sanguineous fiuid in the body cavity. In general, the interior of this bird was more degenerated than that of the undrawn, though the odor of the undrawn, especially the abdominal cavity, was distinctly fecal. The final examination to be reported at this time was made when the chickens had been in storage 610 days. These chickens, as indi- - cated in the table, where they are designated Nos. B. C. 695 and B. C. 696, show a more widespread, and also a more varied, bacterial invasion, than the cold-stored chickens previously examined. Their odor was rancid, the drawn having less odor than the undrawn. The tissues were generally desiccated. The skin was watery in appear- ance. The undrawn chicken was practically free from abrasions. The drawn had over the surface of one thigh an area of torn skin. Both chickens were discolored in the region of the vent. Internally the undrawn chicken showed greenish-yellow fat and flabby heart; dark congested lungs, very watery; a pale yellow liver, very soft; a gizzard which was soft, flabby, and degenerated; intes- tines which were pale, grayish-yellow, watery, and somewhat shiny; a dark spleen and dark kidneys; a reddened bone marrow; body walls which in the region of the liver were deeply stained with bile pig- ment, and in the abdominal cavity itself a considerable quantity of thin, watery, bloody fluid. In the drawn chicken the whole interior of the body cavity was dark in color and very bloody; heart and lungs dark and full of a thin, watery blood. The kidneys had practically the same appearance, all looking like pieces of old leather. This fowl, in general appearance, was not as good as the undrawn. It is of interest to note that the species of organisms in the drawn and the undrawn fowl examined simultaneously have a very strong similarity and in many cases a coincidence. If, as is asserted by some, the bacterial invasion of the fowl in cold storage is due to the migration of the organisms from the intestinal canal into the tissues, one would expect in a drawn fowl, properly prepared, either a lack of organisms or the presence of species different from those found in the undrawn. It is quite possible, however, that the drawing of the fowl may be done in such wise that the organisms of the intestine are’ permitted to gain access to the viscera, in which case they would undoubtedly be found in the tissues. Gas-producing organisms, which have their ultimate habitat in the intestine, have, as will be noticed in the list of species, been isolated in certain cases. How- CHICKENS COLD-STORED UNDER KNOWN CONDITIONS. 55 ever, so widely distributed are these organisms, and so many chances would they have of coming in contact with the flesh of the chicken, that it would be difficult to assert that their presence there is due to migration from the intestine of the drawn fowl or to pollution by its contents in the process of drawing. The pronounced increase in the number of species isolated in the course of the work here reported would indicate one of two things— either that there has been a very decided increase in the number of the organisms originally present in the flesh, enabling the methods, which at the beginning of the experiment were inadequate for their isolation, to meet the conditions present later on; or that the addi- tional species have, in the course of the period of storage, gained access to the flesh. It must be remembered that these chickens were prepared, placed in storage, and treated at every step of the way with a care which is not exercised under practical market conditions. With such pre- cautions as have been taken it would seem scarcely possible that so extensive an invasion as represented by the increase in species found in the later stages of storage could have taken place in the warehouse. It seems probable, though further work is demanded on the subject, that there has been an increase in the number of the organisms in the flesh of the chickens even though the temperature at which they had been kept is far below the freezing point. It has also been noted, in the course of this investigation, that the organisms developing after long periods of cold storage are difficult to classify. It is altogether probable that the low temperatures at which they have been kept for months is, in a measure, responsible for their variation in growth when sown upon the usual culture media. It has been quite impossible to attach a definite name to each organ- ism since there are slight differences between individual colonies which are, however, insufficient to throw them into different groups; and they differ also from the descriptions of the organisms commonly given in the literature devoted to systematic bacteriology. Bacteriological examination of fresh and cold-storage chickens. FRESH CHICKENS. | Cultures. Descrip- | Time of ores No. Date. eee | storage. ae wee Species isolated. | | | | sults.| Ree acs SOO ers | OMNIA Were epee terete Head wound....| + A micrococcus, a rod form. Heart blood..... 0 MVM as cae ste ase 0 Signe Seanedeced 0 Gall bladder ..... 0 iKobolN ie Seoooaeee 0 Bone marrow..-.-| 0 Thigh muscle....| 0 Breast muscle...| 0 74 ae 10-19-06 .|..... GOWER rt Repke as eaae KGHOMC Yi eee o t= ce + Bone marrow....| 0 UE Secrets eo eara ta Thigh muscle....| 0 56 PRELIMINARY COLD STORAGE STUDIES. Bacteriological examination of fresh and cold-storage chickens—Continued. FRESH CHICKENS—Continued. | Cultures. Descrip- Time of No. pate tion. storage. peeans Re- sults. AOKI oe il —=13=0 7-25 al Gnesi eee eee Heart blood....- 0 AVC Sone 0 Spleeneeeses-eee- 0 Bone marrow...-| 0 Breast muscle...| 0 160 Sse) OSI Gailssace GOs 2 Soa hese ae seer Heart blood.....- (0) VCR nae eee 0 Spleene==se eee 0 Bone marrow..--| 0 Breast muscle...| 0 Bac: Be: 10-21-07.|....- dossae inight === 2-> 4|stleart bloodees-= ar | GIVER so soso eee 0 Spleenege. sane. + | erdmey 22a" 2a s- 0 Bone marrow..--| 0 Thigh muscle....| 0 Breast muscle.-| + Ba Orb 947m 23=08ee|seeae Gdoseseee A few hours. .| Heart blood....-| 0 | Divers. soe 0 Spleen: —oeecer ee 0 Kidneyeseeeeae SF Bone marrow...-| 0 Une ee 0 Breast muscle...| 0 Peritoneal fiuid..| 0 Intestine... -2=: + | tilis? COLD-STORAGE CHICKENS. Days D5 eee 1O195065| Daiwa see | ell Oa Heart blood..... + iver saseere ee: | ae Spleenase sees ee 0 Kidneys seas see = tericus. Bone marrow....| 0 Ib bhstees eo Seoeee _ Thigh muscle..... + Peritoneal fluid...) + AD lee ee dOe = | Umadrawine=s|2eee= do-ee- sa Head wound....| + Heart blood..... 0 LVCT ee cee ee 0 Spleen= =e see 0 hKadneyeee- eee a Bone marrow...-| -+ hung ss. 23.2e2e + Breast muscle...) + Peritoneal fluid..) + IMtestines=sscac2 ete J AQ06 Eee SH Gal Denials sal} Alls soe Heart blood..... 0 WGIVER =e ee 0 Bone marrow..-.-| + ciens ? AGU ee Sa Oz Thigh muscle....| 0 Breast muscle...) + | : ciens ? Peritoneal fluid..) + CO ee nl lSace dons) Undrawne.slsecee GOPsa=seee INS CVNG. s a bectlisasssc BeColb sa 6-E 074 SD rawilesess| hoster aes Thigh muscle....} + Breast muscle...) + | A motile bacillus. | M. pyogenes aureus?, B. Species isolated. A bacillus—motile, a mi- crococcus. carneus? A micrococcus. Streptococcus, M. aqua- M. pyog.albus. B. coli var. ?, B. mesen- Aspergillus glaucus, B. coli var. ? B. coli var. ? M. epidermis albus ?, B. fuscus Chester. M. candicans ? Strep. erysipelatos? Large intestine: Pink yeast, Mucor sp. ?, M. aurantiacus, Oidium lactis, B. cinctus Ravy- enel, B. megatherium, B. putrificus ? Small in- testine: Cladosporium-- like fungus. B. fluorescens liquefa- B. fluorescens liquefa- Bact. anaerogenes. Cultures and organisms as in 4296. Rod, not classified. Rod, not classified. MARKET COLD-STORAGE CHICKENS. a7 Bacteriological examination of fresh and cold-storage chickens—Continued. No. ms. ©. 14_- B.C. 561- B.C. 560-|- -. B. C. 696. COLD-STORAGE CHICKENS—Continued. - coli B. fluores- Cultures. Descrip- Time of = 5 a Date. fee storage. ae ee Species isolated. sults. Days CaM sal) Whaisberhyacleacll Bieveassenanese Heart blood....- -f Rod, not classified. IVAN) Ore eer acne 0 Spleennens apace 0 Keidneyeeeeeeecre 0 eM See tes Soke + | Bacterium sp. ? Thigh muscle...-| 0 Breast muscle... Bacterium sp. ?, B. fluo- rescens liquefaciens.a Peritoneal fluid..| + | Bact. aviwm ? Chester. ImMtestine == sees. + | Bact. anaerogenes ? 1O=22-O7F ND rawittererese Ol tence sce eee Heart blood.....- + TGIChA ic cogacseee AF Bone marrow....| 0 Thigh muscle.-..| + Breast muscle...) + |B. liquefaciens, B. var., B. liquefaciens fluo- rescens ?, cens non-liquefaciens. sO. 5) WnachearnnsolossocGlMnqdsocor Heart blood....- 0 : MeV THE A acct sees + M. luteus. Spleen ewe suse + | B. raveneli ? Kenyans eee + | B. cloacae ?, Strep. ery- stpelatos ? Bone marrow....| 0 Thigh muscle....| + | B. fluorescens non-liquefa- ciens, B. germinus Ra- venel ? Breast muscle..-} + M. epidermis albus ?, B. fluorescens ? Intestine........ oF Crop contents...| + | B. subtilis ? 122 (So mel Any ere es | O)l ONeee ents WAV ery is Ser + | A micrococcus-like cul- ture 687 isolated from B. C. 695. Keidneyaeaseosos- ae Bone marrow..--| + Breast muscle..-| + sconO Os col Wiravehehyaats lleosca0 On seasccs Crop contents...) + Strep. enteritis, B. intesti- nalis ?, B. subtilis. Heart blood..... + | B. alcaligenes. GIVE eee + | Micrococcus culture 687. Spleenteeacs se! 0 Kerdmeyer ee seeeee + ; Micrococcus culture 687. Bone marrow..-.-| + Micrococcus culture 687, Sarc. subflava. Thigh muscle....| 0 Breast muscle...) + M. cumulatus ?, micrococ- cus culture 687. ; Peritoneal fluid. .| + Micrococcus culture 687, B. coli. Iimibes tine meee + | Strep. enteritis, B. cloacae. Pericardial fluid.| + | Micrococcus culture 687. an axillary space. IV. MARKET COLD-STORAGE CHICKENS. By M. E. Pennineton, In charge, Food Research Laboratory, Philadelphia. INTRODUCTION. The results to be set forth in the following pages represent a pre- liminary study, on the broadest possible lines, of the action of low temperatures on foodstuffs, more especially those rich in protein, such as meat from various animals, poultry, fish, eggs, ete. Such a study would seem to be a necessity to the well-being of the people 58 PRELIMINARY COLD STORAGE STUDIES. of the United States because of the almost universal use and enor- mous increase within recent years of cold-stored food products of varied kinds and from many sources. The progressive cold-storage warehousemen are also eager for scientifically acquired data, that they may improve their product thereby. From the point of view of pure science the field for investi- gation would seem to be almost limitless and filled to overflowing with information most interesting and valuable to the chemist, bacteriologist, and histologist. The domain of the study is so vast, and the factors which must be considered in the correlation and inter- pretation of results are so numerous and far reaching in their effects, that the question has not been ‘‘ What phase of the problem shall be investigated ?’’, but, rather, “‘Which question can be deferred with least injury to the interpretation of the results already obtained?”’, since all seem to be interdependent and must be taken into account when any individual question is under consideration. In view of the fact that this field of investigation is almost barren of reported results and because of its diversified interests, 1t has seemed better to make a general study of the chemical, bacteriological, and histological conditions prevailing in the case of some standard cold- stored product rather than to follow any one line of scientific work, though such a procedure would have made possible the addition of many details now waiting to be investigated. Poultry in general, and chickens in particular, have been selected as objects for study, not only because they are of enormous economic value, but, being small, highly organized, and easily obtained they would seem to be eminently suited to the present investigation. With the extravagant advocacy of the advantages of cold storage, as claimed by some of those commercially interested on the one side, and the increasing prejudice against cold-stored foods in the minds of the laity on the other, this discussion can not deal. Suffice it to say that such contradictory statements led to the macroscopic exami- nation of chickens which, intended for the general market, had been for varying lengths of time in cold storage, a comparison being made with fresh chickens of like breed and age. The results of such observations would seem to demonstrate that, in appearance, odor, and texture of the flesh, long storage at low temperatures causes a variation from the normal fresh bird, and also that the changes which do occur slowly and requiring long periods are not comparable with those occurring rapidly and at ordinary temperatures. A discussion of the macroscopic changes, particularly those which might be observed by the housewife, has already been published.* A logical sequence of such a study is the microscopic exam- ination of the tissues of such chickens, their chemical analysis, and a « Yearbook of the U. 8. Department of Agriculture, 1907. esi 2 Or MARKET COLD-STORAGE CHICKENS. determination of their bacterial content, on which may depend their fitness for food. The work reported will, therefore, be classified as chemical, bacteriological, and histological; the technique employed and the results obtained for these phases of the subject will be given under their respective headings; and the general discussion of the results, with, as far as is possible at this time, a correlation of the data relating to the changes, will be brought together in a final chapter. CHEMICAL STUDIES. RESUME OF THE LITERATURE ON THE CHEMICAL COMPOSITION OF POULTRY. An examination of the literature relating to the chemical composi- tion of poultry in general, and of chickens in particular, reveals a paucity of analytical data. Such analyses as are given usually relate to the gross composition of the birds, comprising the amount of water, nitrogen, fat, nitrogen-free extractives, and ash. Koénig” has brought together in tabular form analyses made by Stutzer, Moleschott, Kénig, Krauch and Allendorff, and Atwater and Woods, aggregating seven in all. As mean percentage values for the flesh of chickens Kénig gives the following: aS ee ee ae cae ne ee Ce eee ky Mee ay Ge OD Pep tie RCC ere ere pt are ne sn tn eee So Pk eee Sols te oe hee Ble se PREM kw wed eae wR NE ata SSSR Se 2 Sm a Re aa a 4, 5d _ TEDTD INPRO CST NV OND IS ISD: C1 Cl Ea Shh et tna a me a em i ee a 0.75 STS oe DREN SSS A Set Cape har Seats. ee Sera eae em eT a ies, In connection with a study of the meat supply in Tennessee, C. E. Wait© reports analyses of 20 chickens purchased in the open meres in that State. Atwater and Bryant? in their summary of the chemical composi- tion of American food materials, give average data for analyses of chicken and other domestic poultry and of the gizzard, heart, etc. Atwater? later reported a number of additional analyses of poul- try. He examined, as a nutrition study, the carcasses of repre- sentatives of most classes of edible, domesticated birds. His analyses treat not only of the meat, but also of the giblets, the entire fowl, and the meat and giblets together, the latter bemg the truly edible portion. Only the analyses of the meat are available for compari- son with the results obtained in this laboratory and the average data «Chemie der menschlichen Nahrungs- und Genussmittel, 1903. > Calculated by difference. cU.S. Dept. Agr., Office of Experiment Stations, Bul. 53. a@U.S. Dept. Agr., Office of Experiment Stations, Bul. 28. é The ormeion of Poultry, Storrs Agricultural Experiment Station, 15th Annual Report, 1903. 60 PRELIMINARY COLD STORAGE STUDIES. are quoted. On one chicken separate analyses of light and dark meat were made, as will be observed in the table. There has been no attempt, however, to separate or determine the various kinds of nitrogenous bodies which go to make up the sum total of the protein. For further details those interested are referred to the original of Atwater’s valuable communication. Composition of poultry (Atwater). Heat of combus- ES tion per gram. © Description of sample. Water. | Protein. Fat. Per cent.| Per cent.| Per cent.| Per cent. | Calories. 2 cold-storage capons—meat...-...-..---..-.-.----- 55. 8 | 21.6 22.1 1.2 3. 222 ZCHIGKEMS==IM Sayre ee, et ae oe ern a 66.9 22.6 10.1 alt 2. 209 2 chickens—broilers—meat.............-...-.-.-.--- 69. 2 Di 2 8.8 ial 1. 938 2 TOWIS= MO Gee oe ce oe eye eee hie on ib ein aelapen se ee oa ge 58.9 20. 5 19.9 ileal 2. 961 lchicken—Garkmedilaes=-scceeeee eee eee eee eee 70. 1 20. 8 8.2 Ne 1.872 {cchiecken—light mealasseseen- moe oe eee eae 70.3 | 21.9 7.4 ileal 1.845 The literature on the composition of chicken fat is as unsatisfac- tory as that dealing with the muscle. Amthor and Zink® give the following values as The result of their observations on both the fat and the aiee fatty acids: Chickentat=speeific @ravity. <2. xi 2. See es See ee ee ee 0. 9241 Melting point 225 220 oes 2 oe 2 Ae es ev eee 33-40° C. Solidafy ame Pout ed eae ee Se eee 21=27°-.@, Todim‘number: 2 30)set 2308 ae Be Bee eee eee eee eee 66. 7 Sapomriueatiom: memeber ye ps ees ge ce ee eee ee 193.5 Reichert number 2 a. tee es epg a ei eee ee 1.0 Acetyl mum Der e526 yes ae Se Bee ees ee ee 45. 2 Acidity. Giresh). 225 x22 Aes aac le eae chee eee ee ee ee 2 Acidity (several months old)22 0s es eee ee era ee 2.3 Free fatty acid—specifie gravity atila> C23 soe ee es ene 0. 9283 Meeltime pout :< 5. Saye 2 ea a epee ee a ee ee 38-40° C. Nolidihy ine. Pom ti wee see ge mere Bares bene 32-34° C. Wodim mamma erie i Fea SS ee at eee rae gen ee er 64. 6 Saponification numbers... 22. ba: oe ee a er 200. 8 Zaitscheck® gives an analysis of the fat of two chickens, one of which has been fed on corn and the other on corn and milk. His results are as follows: Analysis of fat from corn fed, and milk and corn fed chickens (Zaitscheck). : F No. 1—corn | No. 2—corn © De reuiniineraion. fed. and milk fed. Specific eraviby lab BOK Cis tesa ey eee eee sees a eee reece ens eters 0. 9129 0. 9153 Melt impap oimt Sa oii oe oe eee eee eee ee eee 36° C. 38. 5° C. SKoV io btinvatoyexh OXON a Reet nem ere Ne eee Re Nal Se me Vea skeet ouee Se 17. 4° C. 2225} Melting sp oimbsot ree waittiy ald See eee eee eee eee ee en eee ae BC Be) ie (Ce Soliditivine: pomiisot tree fab bys aot Gl Seer eee eee 34. 5° C. SOROS OE Retractiomrat AO? Cae au Aes ei aie ie OS eeayy eres eee es Sy eee a Src ane EEE 25 52 Saponii cation mlm bere see. as a ae eres eee ee SU Sree soca eet 214 216.8 Nonvolatile dartiiiy: acids! ys ee sores eae a= eee eee te ee ae eer 95. 3 94.8 Lodin-numiber oftats=5 22-25-5505 secs aes es se rayne sees eee eee eee 70. 6 57.6 Lodinmnumberortatiy-acid Si sseeesenee ee eee sae See ese eae ae eae 54.4 45.4 Molatile Tatty eras. ae Ge eye ag eI eg ae pee Ohare dee ea pe een ea 0. 88 0. 88 Kreetatty acidsicalcmlatedasiOleice=ssss--eeeee-eee eee eee eee ee aaa 0. 40 0. 49 aZts. anal. Chem., 1897, 36: 16. b Arch. ges. Physiol., 1903, 98: 614. MARKET COLD-STORAGE CHICKENS. 61 Zaitscheck concludes that the food has influenced the composition of the fat, the fowl on the corn and milk ration showing values nearer those of butter fat than the results found for the chicken fed on corn alone—an observation, which, if borne out by further experimenta- tion, is of considerable physiologic interest. As is to be expected from the small number of analyses recorded, a lack of harmony in technique, and in the use of fowls of unknown _ age, breed, and previous history, there is a wide variation in results. That the analyses of the flesh of animals so highly organized as are chickens should have an exact and fixed composition is scarcely to be expected, even when breed, food, age, and the length of time between killing and examining are the same. That very wide differences do occur when such factors as have been mentioned are not taken into account is not only reasonable, but has been demonstrated by cer- tain analyses given later in this report. SELECTION OF MATERIAL FOR STUDY. Since the work which has been undertaken is a comparison of the ' flesh of chickens kept by the aid of low temperatures for varying lengths of time, it is necessary that there should first be an acquaint- ance with the composition of fresh fowls of known history and of the breeds commonly found in the markets. The species most fre- quently kept for food purposes in the Middle States are the Plymouth Rock (white and barred), White Wyandotte, Rhode Island Red, and Leghorn. The last-named species is especially valuable for egg pro- duction. The birds are too small and the breast muscles are not sufficiently well developed to render them as desirable for market purposes as are the other breeds, but because of their laying quali- ties they are fairly plentiful, and young cocks and old hens are there- - fore killed off and sold for food. As the cold-stored fowls which are to be discussed were taken from regular market stock and are, almost exclusively, of the Plym- outh Rock variety, or its crossbreeds, the discussion of the compo- sition of fresh chickens will be limited accordingly. The birds kept for standards of comparison were fed chiefly on Indian corn, milk, and clean table scraps. They were regularly and abundantly fed, were supplied with pure water, and their runs were placed in an orchard having a southern exposure, this environment insuring rapid growth and tender flesh. For 24 hours previous to killing all food was withheld, but water in plenty was provided. They were killed by puncturing the spinal cord by way of the mouth, and were bled well, dry picked, and the animal heat was quickly extracted. They were kept for from 18 to 24 hours at the tempera- ture of an ordinary refrigerator before the analysis was begun, that being the period required for a frozen chicken to thaw in a house refrigerator. All were undrawn. 62 PRELIMINARY COLD STORAGE STUDIES. The history of the cold-storage chickens is unknown up to the beginning of storage, the date of which is known. They were first | placed in a cold freezer having a temperature of from 4° to 6° F. until solidly frozen, which requires about 48 hours; then they were transferred to a temperature of approximately 13° F. All of the market cold-storage chickens reported at this time have been treated similarly. They were removed from the warehouse about 24 hours before the analysis was begun and were allowed to thaw at the tem- perature of an tce box. Occasionally, especially in the case of large birds, there would be enough ice in the tissues when the examina- tion was begun to cause the knife to grit, but usually they were simply very cold to the touch. One chicken only—-No. 69, in storage two years—was thawed in the usual commercial fashion, namely, by soaking in cold water. Since such treatment prevents any accurate idea of the bacterial content of the flesh itself, and as a bacterial, a histological, and a chemical examination of each fowl was desired, dry thawing was adopted. Such a procedure gave a greater proportion of solid mat- ter in the flesh than was obtained in either the fresh or the soaked chickens, because of the marked drying of the tissues while in storage. CHEMICAL TECHNIQUE. The methods of chemical analysis employed were, as nearly as possible, those officially recommended and commonly used in flesh analysis. A brief outline of the procedure followed is given. The chemical analyses were made by J. S. Hepburn and John I. Burrell. PREPARATION OF SAMPLE. When the birds were large enough, only one was used for analysis. When small, two were used, care being taken to thoroughly mix the tissue. The flesh of the chicken was prepared for analysis by separating it from the bones and the skin, removing as completely as possible the fat between the muscles and also the large tendons, which in the leg and wing muscles of the chicken are easily stripped loose from the meat. Their absence is a factor too small to influence analyt- ical results, but their presence renders the grinding and accurate sampling of the meat more difficult. The muscles of the thighs and legs were classed as dark meat, of the breast and upper wings as light meat. The dark and light portions were put separately through a meat grinder until the particles were very finely divided, which necessitated ordinarily three grindings. It was found undesirable to erind the flesh too finely, since the shaking required for the extrac- tion of the soluble material caused an emulsion which 1t was practi- cally impossible to filter. MARKET COLD-STORAGE CHICKENS. 63 The skin, with the adhering subcutaneous fat, was mixed with the fat separated from the viscera and the abdominal cavity and that stripped off of the muscles, ground in the meat cutter until fine, and used for the study of the fat of the fowl. MuscLeE DETERMINATIONS. The general analysis of the muscle was based on the determina- tion of water, fat, ash, total nitrogen, and total solids. The nitrog- enous portion was further investigated for the nitrogen soluble in water, and in this aqueous extract was determined the coagulable ni- trogen, albumose nitrogen, and amido acid nitrogen, the peptone nitro- gen and the nitrogen insoluble in water being found by difference. For the determination of the total solids, water, fat, and ash in the muscle, a homogeneous well-mixed sample of the meat was weighed in a tightly stoppered weighing bottle, and portions, of approximately 10 grams each, were transferred to appropriate con- tainers, as desired. TOTAL SOLIDS AND WATER. For the total solids and water a 10-gram portion was dried at 100° C. in a tared lead bottle cap from 2 to 3 inches in diameter. It being practically impossible to dry a sample of such material as chicken meat to constant weight, because of the oxidation of the substance after the removal of the last traces of water, the sample was reweighed at short intervals and a gain in weight taken as an indication of the absence of moisture. This gain was ordinarily not more than a few tenths of a milligram. Occasionally, however, it was several milligrams. FAT. The quantity of fat in the muscle material itself was found by extracting the dry residue from the water and total solids determi- nation with Squibb’s ether. This was readily accomplished by cut- ting up the lead cap, with its contents, into small pieces and placing the mixture in the thimble of a Knorr extractor, which was then, with the requisite amount of ether, heated for 16 hours on an elec- tric heater. When the extraction was completed, the ether was evaporated and the fat dried at 100° C. until the moisture was removed, which was indicated by a slight gain in weight, as in the case of the total solids. The weighings here were at intervals of from 2 to 3 hours. ASH. For the determination of the amount of ash two or three grams of the sample were dried in a platinum crucible, then heated in a muffle until completely charred, allowed to cool, and extracted sev- 64 PRELIMINARY COLD STORAGE STUDIES. f eral times with hot water. The carbon insoluble in water, and the small ashless filter paper required for the extraction, were returned to the crucible and the ignition repeated, using ammonium nitrate, — if necessary, to oxidize the last particles of carbon. To the grayish mass the aqueous extract was then added, dried on a water bath, and ignited in a muffle until the ash was clean. NITROGENOUS CONSTITUENTS. The study of the nitrogenous constituents was carried out as fol- lows: The total nitrogen of the two kinds of muscle was determined by the Gunning modification of the Kjeldahl method. For the study of the water soluble nitrogen a portion of the finely divided red or white meat, weighing 60 grams, was put into a tall slender bottle of 500 ce capacity, constructed to fit a centrifuge capable of carrying 1 liter of material; 300 ce of water were added and the flask gently shaken for 15 minutes. The movement was merely sufficient to keep the particles of meat in motion and the composition of the extract homogeneous. Forcible shaking caused an emulsion to form, as did the very fine grinding of the tissue. After shaking for the required length of time the flask was rotated in an electric centrifuge for 20 minutes, which caused the heavier particles to settle in a com- pact mass and permitted the decantation of the supernatant liquid, which was then filtered through paper. The extraction, as outlined, was repeated with portions of 300 cc of water until the filtrate was — practically proteid free, as indicated by the biuret reaction. The — attainment of this result required ordinarily a volume of 1,500 cc to — 2,500 cc. To guard against bacterial decomposition thymol was — added both to the flesh and to the extract, and to inhibit, so far as — possible, the action of the naturally occurring enzymes of the meat, | the solution and the meat itself were kept cold, ice being used when | necessary. The extraction of the white meat was a much simpler operation than the extraction of the dark meat. The latter did not settle as compactly in centrifuging, filtered more slowly, and persisted in show- ‘ing a distinct biuret reaction for a considerable time after the white meat was free of water-soluble proteid. In fact, certain fowls, more especially those which have been in cold storage for long periods of time, never showed a red meat entirely free from water-soluble nitro- gen. In such cases the question of the error due to long manipula- tion and enzyme action, involving a rise in the actual quantity, had to be considered. It was found by experiment that after long extrac- tion of such tissue a point was reached when a very faint biuret reac- tion, which did not apparently diminish, persisted indefinitely. Such extractions were halted after about 26 hours, it being believed that a greater error would result in the gain of what had been originally MARKET COLD-STORAGE CHICKENS. 65 insoluble material than in the loss of the preformed water-soluble nitrogen. The total extract of the muscle was made up to a definite volume and neutralized to litmus paper with tenth-normal sodium hydrate. Duplicate portions of 100 cc each were transferred to beakers, evap- orated to very small bulk (about 10 cc), and transferred with the aid of sulphuric acid to Kjeldahl flasks for the determination of total nitrogen soluble in water. Duplicate portions of 350 cc were heated on the water bath in order to determine the amount of coagulable nitrogen present. It was found better for the complete separation of the proteid to evaporate before filtering to about 100 cc. The coagu- lum of the white meat separated more easily and in a much more com- pact mass than did the coagulum of the dark meat. Both were diffi- cult to wash free of proteid material. The coagulum and the filter paper were transferred to a Kjeldahl flask for the determination of the amount of nitrogen. The filtrate from the coagulum, having been considerably increased in volume by the washings, was reduced to 100 cc. Fifty cubic centimeters of this were taken for the determination of albumose, which was made according to the method of Bémer.* Concentrate the liquid to 30 ce and allow it to cool; add 1 cc of 50 per cent sulphuric acid, saturate with zinc sulphate, warm on a water bath with stirring until clear, and allow to stand 12 hours for the separation and deposition of the precipitate. Filter cold and wash with a saturated solution of zinc sulphate acidified with sulphuric acid. The determination of the nitrogen is made as usual. In the other 50 ce portion of the filtrate from the coagulated pro- teid the amido acids were determined according to the method of Bigelow and Cook,? which, briefly stated, consists in the addition of 15 erams of sodium chlorid to 50 cc of the amido acid containing extract, chilling thoroughly in an ice box, then adding 30 cc of a 24 per cent solution of tannic acid, making the volume up to 100 cc, and allowing the whole to stand at the temperature of an ice box for 24 hours, after which the precipitate is filtered off. The solution is kept cold mean- while, and the nitrogen in 50 cc of the filtrate is determined according to the usual Kjeldhal procedure, except that the addition of potas- sium sulphate is unnecessary, because of the large amount of sodium chlorid present, and care must be taken during the early part of the oxidation that the escaping hydrochloric acid does not cause the mate- rial to foam out of the flask. Since tannic acid invariably contains a certain amount of nitrogen, a blank must be run with every experiment. aZts. anal. Chem., 1895, 34: 562. bJ. Amer. Chem. Soc., 1906, 28: 1485. 49078—Bull. 115—08——5 66 ; PRELIMINARY COLD STORAGE STUDIES. Fat DETERMINATIONS. The study of the fat consisted in the determination of the iodin — number, the saponification number, the acid value, ester value, the Hehner number, and the index of refraction. | The fat of the fowl, prepared as previously described (page 63), was | heated for 2 days in the thimble of a Soxhlet extractor, with a petro- leum ether having a boiling point of between 40° and 60°C. If any solid particles separated after the extraction the solution of the fat | was filtered through paper, and the ether was then distilled off. Dry- | ing the fat, with the small amount of adhering tissue, before extract- ing, was not practiced. The quantity of tissue was very small, because the mechanical separation was carefully made, and it was deemed better to risk the mixture of a certain amount of nitrogenous extractives with the fat than to subject it to the splitting produced by the action of air and heat. It is proposed to make a study of these two procedures to determine their relative accuracy. It was found exceedingly difficult to remove the last traces of petroleum ether without altering, to a greater or less extent, the composition of the fat itself. The result was accomplished best by allowing the fat, freed as far as possible from petroleum ether by means of distillation, to stand in a vacuum desiccator over calcium chlorid freshly ignited and acid free. It was found that sulphuric acid, in a vacuum desiccator, very materially altered the value of the fat, probably owing to the absorp- tion of a certain amount of sulphuric acid vapor. Though it is generally agreed among bacteriologists that pure dry fat is not a culture medium for bacteria, the precaution was never- theless taken to preserve the samples of fat, even for the short interval between the distillation of the petroleum ether and their further anal- ysis, in sterile flasks, plugged with sterile cotton, and the transfer of the material when necessary was made with sterile pipettes. Because of the action of light and air on pure fat the analysis of the samples was conducted with all possible haste. The iodin number was determined according to the official Hanus method.“ For the saponification number the official method was also followed, and the Hehner number was determined as officially recommended.“ The acids, however, were dried at 100° C. until a gain in -weight occurred, when the minimum weight was taken as expressing the quantity of acid. | For the acid value 4 or 5 grams of the fat were weighed in a 250 ce Erlenmeyer flask; 100 cc of a 15 per cent sodium chlorid solution were added, and the flask was immersed in hot water until the fat was liquefied. The free acid was then titrated with tenth-normal sodium hydroxid, using phenolphthalein as an indicator. Vigorous «U.S. Dept. Agr., Bureau of Chemistry, Bulletin 107, pp. 136-139. t ) MARKET COLD-STORAGE CHICKENS. 67 and frequent shaking is necessary to determine the permanence of the delicate but distinct pink which indicates the end of the titration. For convenience of expression the acid value has been calculated also as oleic acid, which is readily accomplished, since the acid value is expressed as fuillienanis of potassium hydrate. The difference between the acid ie and the saponification number represents the ester value. The refractive index? was determined independently by three or four observers, and the mean of all of the observationstaken. AnAbbe refractometer was used. The readings were taken at a temperature between 30° and 40° C. and corrected to 35° C. by the use of the factor 0.000365 for 1° C. || ANALYSES OF FRESH CHICKENS. The values obtained in the chemical study of fresh chickens are given in Tables A to D. In Table B the comparatively low acid value shown by this particular fat is striking, and in Table C the relatively large amount of protein in the light meat as compared with that in the dark and its greater solubility in water are to be noted. 3 Throughout all the analyses there are decided differences between the individuals themselves. Whether such variations can be traced to definite conditions it will take many more analyses and much study to determine. It is believed, however, that the data on these fresh chickens are sufficient to show that certain differences do exist between them and chickens preserved for varying lengths of time in cold storage. (A) Percentage composition of fresh chicken muscle. | Sum of a Protein, Kreatin : Sample. ment Water.| Fat.|Ash.] (NX | (Nx | On o8 oniche : 6.25). | 3.11). : s de termined. Line : lsLight...| 75.50 | 0.49 | 1.17] 23. 46 1.10 | 24.50 97. 92 No. 66. Plymouth Rock broiler.---4 Hark) !_| 71.75 | 2.40 | 1.21 | 21.40 0.827 | 28.25 101. 33 No. 68. Plymouth Rock young NOUSvEI Ol DLOlenee =e has eee Mightase| dowdon| Os lz Dark...| 75.86 | 1.38 .33 | 21.84 1.01 24. 27 100. 08 49) 21.07 0. 64 24. 14 100. 44 24 | 22.52 0.920 | 24.70 100. 92 .18 | 20.69 0.743 | 25. 52 99597 26 | 23. 50 1. 01 26. 44 100. 31 — i ItieAsie 54] 765330 || Oil Dark...| 74.48 | 2.88 Pights 73. 56 | 0.98 et et No. 73. Plymouth Rock roaster. . at No. 78. Rhode Island Red roaster. { _— jt ies Dark . 73.02 | 2.99 35 | 23.13 0. 64 26. 99 101.13 No. 86. Rhode Island Red large (tiene. Pelee OORT ONoan | le21, | 2195 1. 02 24. 99 99.13 “TEC eS eae ee ee Maken |e (onOs |e 2elonl loan el Oid 0.796 | 24.06 99. 78 a2U.S. Dept. Agr., Bureau of Chemistry, Bulletin 107, p. 131. 68 PRELIMINARY COLD STORAGE STUDIES. (B) Analysis of fat of fresh chicken. Plymouth | Plymouth | Plymouth | Rhode Rock Rock Rock Island Red Determinations. broiler, broiler, broiler, roaster, Average. | No. 66. No. 68. | No. 73. No. 78. Todin TWIMbeEsss¢ 9e3e eee eee | 62.6 a2 62.7 61-1 64.4 saponification numbers. s=2..—_ a ese | 182. 2 176.7 190. 2 176.8 181.4 ACI Vale Se see aie Sea ee eee an ee 0.5 1.5 0.8 0.5 0.7 Hester vValuiGw. S222 cece ee ee eee 181.7 175. 2 189. 4 176.3 180. 6 Per cent of free acid calculated as oleic. -..- | 0.25 0.76 | 0. 40 0. 25 0.41 - Hehner number 22a. ss ees 86.8 84. 25 88.7 85. 79 86. 36 (C) Percentage of nitrogen in muscle of fresh chicken. LIGHT MEAT. Determinations. | No. 66. | No. 68. | No. 73. | No. 78. | No. 86. | Average. Total nitrogenke ss. aee nen 2a ee ee | 4.11 3. 82 3. 90 4. 07 3. 84 3.94 > Total nitrogen in aqueous extract ......- {, Roe eke ee eee mae i 23.91 Coagulable nitrogen in aqueous extract -- { eS None | coe eee aoe \ 10.42 } . 0. 0269 .0290 | 0.0181 0. 01 0. 0392 | = Albumose nitrogen...-..--+.-----+--+-+++- 0.652 | 0.775 | ores | O:865) eae i 0.675 — | | Amido acid nitrogen... 22-22-52 cece Raa pa = aa can 2 a2 } 8.27 5 ee e . e | Peptone nitrogen (by difference) ........- ee Se ener ee ie re \ 4.19 Nitrogen insoluble in water (by differ- { 3.19 2. 86 2.90 Sal5 2.917 \ 76.09 ONCE) oe aes A ee 77.60 74.8 74.5! 77.5 76.00 : DARK MEAT. Motal mitrosen<4 pasos eee 3.69 | 3.58 3.55 | 3.91 3. 42 3. 63 Total nitrogen in aqueous extract-.----- i eae pee Ree ees | “eee \ 17.73 Coagulable nitrogen in aqueous extract. - { aaa | ORS oe ae | Z ee \ 8.05 : ‘f 0.0235 | 0.0205) 0.0112) 0.0103} 0.0215 Albumose nitrogen. ....-.-.---+-+-+++--+- 0.637 | 0.573 | 0.316 | 0.263 | 0.628 \ 0.483 | 208 | 0.2 : Amido acid nitrogen ...+.2++0.2+eesecs7 { an (cc) és | 5.33 | Fas 668 © | | Peptone nitrogen (by difference) .--.-.-.... { Bons Ricans aoee ee 0. se \ 2.39 Nitrogen insoluble in water (by differ- 3. 03 2. 88 2.91 Sy eo) Ei I 82.27 ence) = sola a ce wla = 6 wiele =a a ale sa me's a)e sam =\elolera te 82.00 80.5 82.1 85.5 81.3 | cote a All figures in bold-faced type are percentages based on total nitrogen. MARKET COLD-STORAGE CHICKENS. 69 3 (D) Nitrogenous constituents of fresh chickens (calculated on water- ash- fat-free basis). LIGHT MEAT. | Determinations. No. 66. | No. 68. | No. 73. | No. 78. | No. 86. | Average. Per cent.| Per cent. | Per cent. | Per cent. | Per cent. | Per cent. On aeMUPLOSSMn faecal c ccs eases ot eee 19. 74 16. 77 16. 95 16. 80 16. 38 17. 32 Total nitrogen in aqueous extract.-.....- 4. 43 4.19 4. 33 3. 81 3. 94 4.14 | Coagulable nitrogen in aqueous extract. - 1. 626 1.79 2. 32 1. 66 1. 58 NS 206 EN DUM OSE MITTOLEN!. ee SPY TRS UF RE, ty a "i \ \ pus) Se mF Ae PP os x > os 12 , a = i PER ——— _ ee on Bes ai aw Daye a ifs Re 6 Va - - . ; eal , 4 Bee le evan : 4. 4 » ; 2 \ - Ak, noe alS NG Fic. 2,—TRANSVERSE SECTION. SOAKED BREAST MUSCLE OF CHICKEN IN Cotp StToRAGE 2 YEARS. [Zeiss: 16mm. objective, apochromatic; No. 12 ocular, compensating. ] Bul. 115, Bureau of Chemistry, U. S. Dept. of Asriculture. Fic. 1.—LoneitupINAL Section SHowING VACUOLATED DEGENERATION. (Zeiss: 4mm. objective, apochromatic; No. 8 ocular, compensating. | Fig 2.—LoneitupDINAL SECTION SHOWING VARIED AND EXTENSIVE DEGENERATION. BREAST MUSCLE OF CHICKEN IN CoLD STORAGE 4 Years. [Zeiss: 16mm. objective, apochromatic: No. 12 ocular, compensating. MARKET COLD-STORAGE CHICKENS. 85 ical appearance is more than explained by the struct of the tissue as revealed under the microscope. Plates XII and XIII show the character an the intestine of a fresh chicken and also of a chicken in cold storage for six months: It will be observed that the muscular coats of the cold-stored chickens have quite lost their characteristic structure. Apparently they con- sist almost exclusively of the sheath of the fibers which, if they con- tain muscle substance in any quantity, have it so altered chemically that standard muscle-staining dyes do not affect it. What remains of the individual fibers is a mass, loosely put together, which would offer but little resistance to the migration of bacteria. So far as the cellular portions of the intestine are concerned they are reduced to a minimum. Either the intestines are obliterated or the cells are so dried and disintegrated that they are mere fragments. Though the cell, as a whole, is in bad condition, the nucleus has apparently offered greater resistance to the corroding action than has the cytoplasm of the cell. GENERAL DISCUSSION. Bacteria and enzymes are most directly concerned in the question of flesh decomposition, the effect of air, light, temperature, humidity, and such conditions as are ordinarily considered responsible for altera- tions in organized tissues being in reality factors influencing the growth or activity of one or the other, or both, of the agencies stated. The investigations of flesh decomposition which have been reported have been conducted usually at room temperatures or at body heat. - Such studies at temperatures near 0° C. are almost entirely wanting, probably because of the general idea that enzymic and _ bacterial activities cease at or about zero. RESISTANCE OF BACTERIA TO LOW TEMPERATURES. The resistance of bacteria to low temperatures is well known, and a number of investigators have subjected various species of organ- isms to almost the lowest limit of modern cold. In 1884 Pictet and Young % exposed B. anthracis, B. subtilis, Micrococcus leuteus, the bacillus of symptomatic anthrax, beer yeasts, and smallpox vaccine to temperatures varying from —70° C. to —130° C. and for periods of from 20 to 108 hours. The bacteria proper were not destroyed; the yeasts were intact microscopically but their functions were lost, as was also the activity of the smallpox vaccine. Macfayden ° exposed a number of bacteria, both benign and pathogenic, to the temperature of liquid air (— 190° C.), first for a period of 24 hours and @ Compt. rend., Jan. to June, 1884, p. 747. b Lancet, 1900, 7 [849]: 11380. 86 PRELIMINARY COLD STORAGE STUDIES. then for 7 days. Buchner’s zymase was also tested and survived for 20 hours. Some photogenic bacteria exposed to this extreme cold promptly ceased to emit ight, but thawing soon caused a resumption of activities. Macfayden says: “It is a remarkable fact that, notwithstanding the enormous mechanical strain to which the organisms must have been exposed, a strain far exceeding in amount any capable of being produced hitherto by direct mechanical means, not the slightest structural alteration could be detected.”’ There was no impairment, according to the observations of this author, in the vitality of the organisms. JBelli® has also subjected the bacteria of chicken cholera and anthrax to the action of the temperature of liquid air, without altering their properties. DEVELOPMENT OF BACTERIA AT LOW TEMPERATURES. At temperatures approaching the zero point not only viability but fertility has been proven, and the literature shows that quite a num- ber of investigations have been conducted, and the field is wider than would at first sight appear. Forster,’ for example, finds that certain photogenic bacteria grow at 0° C. and that organisms capable of developing at this temperature are to be found in earth, milk, flesh, spring water, etc. He also states that the plague bacillus is capable of growing at zero. He determines, too, the rate of growth — in an ice calorimeter, and finds that from 10 to 12 days are usually sufficient. Discussing the question of bacterial growth m_ the German “‘ Kithlhaus,” Forster’ concludes that the changes induced are slower than at ordinary temperatures, but are not stopped nor essen- tially altered. S. Schmidt-Nielsen? found five species of bacteria which developed at zero in the Strassburg water supply, from 10 to 40 days being required for appreciable growth. He found, also, an actinomyces which required 80 days for development. These organisms are Bact. fluorescens nonliquefaciens, B. granu- losum, B. paracoli gasoformans anindolicum, B. tarde fluorescens, and B. radiatum. Fischer’ and Havemann/? have also studied organ- isms which are capable of making a growth at zero, the former iso- lating fourteen species from various sources and the latter obtaming — a Riforma Medica, 1901. b Uber einige Eigenschaften leuchtender Bakterien, Zentrbl. Bakt. Paras., 1892, 12: 431. c Uber die Entwickelung von Bakterien bei niederen Temperaturen, ibid., p.431. 4 Uber das Vorkommen psychrophiler Bakterien, Zentrbl. Bakt. Paras., 1902, 9: 145. € Bakterien Wachstum bei 0°, etc., Zentrbl. Bakt. Paras., 4: 89. fInaug. Diss. Rostock, 1894, Uber das Wachstum von Mikro-organismen bei Eis- schranktemperatur. Bul. 115, Bureau of Chemistry, U. S. Dept. of Agriculture. SMALL INTESTINE OF FRESH CHICKEN. [Zeiss: 4mm. objective, apochromatic; No. 6 ocular, compensating. | PLate XII. Bul. 115, Bureau of Chemistry, U. S. Dept. of Agriculture. PLATE XIll. ra AHL NN ’ AYA SMALL INTESTINE OF CHICKEN SIX MONTHS IN COLD STORAGE. [Zeiss: 4mm. objective, apochromatic; No. 6 ocular, compensating. | MARKET COLD-STORAGE CHICKENS. 87 some from sewage. Miiller “ has isolated 36 species of fungi that will grow at 0° C. from such sources as flesh of fish and cattle, intestinal contents of fish, vegetables, grain, and garden earth. Ten of them were found in flesh and nine in earth. Not only are such organisms widely distributed, but very pro- nounced results in nature have been credited to them. Schmelk ? believes the green color of the Norwegian glaciers to be due to the presence of B. fluorescens liquefaciens, an organism which he finds in such ice, and which develops rapidly at zero. Glage ¢ ascribes con- siderable importance in the maturation of meat to what he terms ‘‘aroma-producing bacteria,’”’ organisms which develop in the German “ Kiihlhaus,’”’ where the temperature is usually from +2 to +5° C., and which he finds infecting not only the rooms, but the carts, trucks, and, in fact, the entire slaughterhouse. Their growth at +2° C. is rapid; at 37° C. it is very slow. That naturally occurring bacteria will develop in milk kept at —1.67° C., with a fair degree of rapidity and in numbers which finally reach several billions per cubic centimeter, has been demonstrated by Pennington.? Certain species of organisms were found to outrun others so decidedly that they were finally present in almost pure cul- ture. Among such were most commonly found B. solitarius Rave- nel, Bact. aerogenes, B. coli, and B. formosus. However, a number of other species also grew below 0° C., as indicated by plates made and incubated at the temperature at which the milk was kept, namely, —1.67° C. These bacteria developed in. spite of the fact that the milk was a semisolid mass of crystals, so solid, indeed, that it had to be dipped out of the container. A multiplication of organisms in ice cream when kept in a frozen condition, either by packing in ice-salt mixture, which gives a temper- ature of about —17° C., or in a cold-storage warehouse where the tem- perature varied from — 18° to — 23°C., 1s reported by Wiley.¢ Appar- ently there is a series of curves of growth, varying both in periods of time and in intensity for different samples, and, in all probability, depending on the rise and development of a succession of species. It is commonly stated that pathogenic organisms do not develop at zero, and, broadly speaking, such is usually the case. That it can not be applied as a sweeping assertion is indicated by the statement of Forster, previously cited, that the bacillus of the plague does grow at @ Arch. Hyg., 1903, 47: 127-193. 6 Kine Gletscherbakterie, Zentrbl. Bakt. Paras., 4: 545. ¢ Zts. Fleisch- Milchhygiene, 1900-1901, p. 131. d Bacterial Growth and Chemical Changes in Milk Kept at Low Temperatures, J. Biol. Chem., 1908, 4: 353. € Milk and Its Relation to the Public Health, by various authors, Hygienic Labora- tory, U. S. Public Health and Marine-Hospital Service, 1908, Bul. 41, p. 257. 88 PRELIMINARY COLD STORAGE STUDIES. zero. Conradi and Vogt” have found that B. proteus fluorescens will grow at zero, and B. Fischer? found a pus-forming coma bacillus which flourishes at this temperature. INHIBITORY ACTION OF LOW TEMPERATURES ON BACTERIA. ' The reverse of the question of life and reproduction at low tempera- tures may be found in those investigations dealing with the destruc- tion or inhibition of bacteria by cold. Repeated attempts to induce certain pathogenic organisms to grow at temperatures even somewhat above zero have failed, as did those of Brehme,° who worked on chol- era and typhoid, and of Dieudonne,’ who sought through successive lowering of the temperature by 5° stages to cultivate an anthrax bacil- lus resistant to cold; +10° C., however, was the limit of growth for this organism. In relation to the self-purification of ice from bacteria Pruddené was one of the earliest workers. He concluded that 51 days were suffi- cient to kill B. prodigiosus; Stap. pyogenes aureus was reduced to one-fifth of its origmal number in 66 days, and B. typhosus was greatly reduced in 103 days. Alternate freezing and thawing is more effective as a killing agent than a single freezing of longer duration. Prudden states that the act of freezing itself either kills or reduces vitality, an opinion in which other investigators, Macfayden, for example, do not concur. Undoubtedly, however, according to the work of Sedg- wick and Winslow,/ continued effects of freezing temperatures, at least so far as organisms in water are concerned, tend to greatly reduce their numbers or even to kill them outright. These authors, however, lay particular stress on the time factor, believing that ice from polluted streams should be stored for months to permit the cold to thoroughly act on the organisms present. Smith and Swingle? believe that the effect of very low temperatures is overestimated and that the cold produced by ice and salt mixture, about —17° C., is as efficacious as liquid air. They hold that the critical point for the generality of organisms lies at or near 0° C., but a Bin Beitrag zur Atiologie der Weil’schen Krankheit, Zts. Hyg. Infekt., 30: 287. b Deutsche med. Woch., 1893, No. 25. c Uber die Widerstandsfahigkeit der Cholera Vibrionen und Typhusbacillen gegen niedere Temperaturen, Archiv. Hyg., 40: 320. d Beitrage zur Kenntnis der Anpassungsfaihigkeit der Bakterien an urspriinglich ungiinstige Temperaturverhaltnisse, Arb. aus dem Kaiserl. Gesundheitsamte, 9: 492. € Bacteria in Ice and their Relations to Disease, with Special Reference to the Ice Supply of New York City, Medical Record, 1887, 31: 341. / Experiments on the Effect of Freezing and Other Low Temperatures upon the Vitality of the Bacillus of Typhoid Fever, with Considerations Regarding Ice as a Vehicle of Infectious Disease, Mem. Amer. Acad. Arts and Sci., 1902, 72 (5). gDer Einfluss des Gefrieren auf Bakterien, Zentrbl. Bakt., I Abt. Ref., 1905-6, 37: 357. MARKET COLD-STORAGE CHICKENS. 89 single individuals may withstand unfavorable conditions. Repeated freezing and thawing will completely exterminate some varieties, cer- tain individuals of which will survive a single freezing and thawing. The interesting observation is also made that spore formers are more resistant than are vegetative cells. From the foregoing summary it would appear that a considerable number of bacteria can, when the food supply is favorable, reproduce at 0° C. or even below that temperature, though growth is greatly retarded, and that freezing, even though prolonged, does not of neces- sity indicate sterility. CHEMICAL CHANGES PRODUCED BY BACTERIA AT LOW TEMPERA- TURES. For the purposes of this investigation the proof of the viability or even of the reproduction of bacteria at low temperatures does not suf- fice, since the changes taking place in certain foodstuffs kept in a frozen condition are under consideration, which changes are funda- mentally chemical. It becomes necessary, then, to investigate the ability of bacteria, when maintained at low temperatures, to cause changes in such compounds as proteins, carbohydrates, and fats, it being commonly held that even if organisms should multiply under such conditions the results of their life processes would not correspond with those taking place at the optimum growth temperature. Miller” has studied the evolution of carbon dioxid, hydrogen sul- phid, the breaking down of protein, and such chemical changes as commonly accompany the growth of bacteria when the germs were maintained at a temperature of zero. He finds that for organisms growing at zero ch chemical indications of activity are not wanting, but that the formation of the various decomposition products is oreatly retarded and their production is very slow. Such results are quite in accord with another observation of Miiller’s, namely, that the life cycle of one of the organisms isolated, B. fluorescens liquefaciens, at 0° C., is twenty-four times as long as is the cycle at 25° C. ‘EFFECT OF LOW TEMPERATURES ON ENZYMES. On this most important phase of bacterial growth and development there is almost a total lack of recorded observations, those changes which have been traced chemically in flesh foods chilled or frozen being ascribed to enzyme action exclusively. Undoubtedly enzymes play an important réle in these decompositions, but it is quite possible that they are not the sole agency. Fermentation changes due to bacteria a Uber das Wachstum und die Lebenstitigkeit von Bakterien, sowie den Ablautf fermentativer Prozesse bei niederer Temperatur unter spezieller Beriicksichtigung des Fleisches als Nahrungsmittel, Archiv. Hyg., 1903, 47: 127. 90 PRELIMINARY COLD STORAGE STUDIES. in milk kept at —1.6° C. have been observed by Pennington.“ In these experiments the acidity of the milk, which is due to bacterial activity, increased enormously. There was also observed a very pro- nounced hydrolysis of the casein, but such a degradation might be ascribed either to the natural enzymes of the milk or to those produced by bacteria or to a combination of the two factors. That enzymic activity is not stopped at 0° C. or even below it seems to be generally accepted, and neither are the enzymes destroyed by cold. Macfayden® found that Buchner’s zymase withstood for 24 hours the temperature of liquid air. Very recently Kovehoff? found that freezing for 24 hours did not destroy the proteolytic enzyme in wheat, peas, and the tissue of Vicia faba. After thawing, the enzyme caused a decrease in protein nitrogen, the nonprotein nitrogen in one experiment reaching 48.7 per cent. Though able to survive low temperatures, the common enzymes are greatly hindered, and the usual course of action of some of them is so altered that their characteristic expression of it is quite lost. For instance, Miller,? experimenting with the action of rennin on milk, found that curding did not take place at 0° C., but, on the other hand, the enzyme still functioned to some extent, because, when brought to 40° C., the curding time was greatly accelerated. The following table illustrates this point: Coagulation at 40° C. ai 5 ee milk at 0° C. + lee of rennin, kept at 0° C. for— Cran Minutes. | Seconds. (OURO ANU OLD R:\c eee a en aoe Ae Re Gee War elnino ee aes ee aise 2 00 (DVM GOS = oo rh nw ee ae ee Es geen Sop Sa 1 50 OOMITTUTE Se So ee ee rd oo pc sk a Le ees Cente Bee ee eres gee 1 45 LOS MINAS se sR ee ay eta a a ane eg SOO 1 10 ADO RMIT EOS 2 Sos a ee ek Seg eg ne Re en 7 | cane ee 55 PSO MMINUGES 2 Fat <= So ce es ee a er gn OV ae | ra 45 DATO UES wpe oe ee IS eS I SS I ee an Lee cain | ae 45 ASSHOUTSE 2 8 304 ges A ee See aa eee ae Pe Stee Se IE oe ee sb Senden 45 QGINOUTS oe eS SIR SE re nie Ae OE Ce eng en | EE RRP 50 After 96 hours, at 0° C., the coagulation occurred as a fine flocking which made its recognition difficult. If other enzymes behave simi- larly it would tend to throw light on many obscure flesh decomposi- tions which take place after cold storage has been maintained for long . periods. Some milk enzymes, however, are not only partly but wholly able to function, as is indicated by the fact that galactase acts in the ripening of cheese at low temperatures, and also Jansen’s statement, that this enzyme does not act more rapidly at 40° C. than at ordinary temperatures. The optimum temperature for the action of the common enzymes lies not far from 40° C. However, it is not yet definitely proven that, @ Loc. cit. 6 Ber. d. botan. Ges., 25: 473. i #! Es LJ MARKET COLD-STORAGE CHICKENS. 9] for certain animals, digestion does not proceed better at temperatures much below this. Miller * compared the action of pepsin from the stomach of the pike at 0° and at 24° C. and found that it took seven times as long to accomplish a given amount of work at the former as at the latter temperature, the pepsin being very active at 24° C. Flaum,’ working on the ordinary functions of the stomach, states that undoubtedly activity does not cease at zero. Fick and Murisier °¢ state that the gastric juice of the pike can digest proteid at 0° C., and Hoppe-Seyler,? who studied the same fish, concluded that not only did digestion proceed at zero, but it was most rapid at 20° C., and quite active at 15°C. Krukenberg, on the other hand, finds just the oppo- site—that 40° C. is the more favorable temperature, and his work is confirmed by Luchhau./ Apparently the resistance to cold is not limited to proteolytic enzymes, since Kastle and Loevenhart 9 have noted that lipase, though splitting fat most rapidly at 40° C., is still active at —10° C., but its action is much slower. At 40°C., for example, 11.29 per cent of fat was hydrolysed, while at — 10° C. only 0.70 per cent was changed. THE ACTION ON FLESH OF BACTERIA AND ENZYMES AT LOW TEMPERA- TURES. REVIEW OF THE LITERATURE. A review of the chemical, bacteriological, and histological data ~ recorded in connection with the analysis of the market cold-storage chickens made in this laboratory indicates that undoubtedly a certain amount of chemical change, and depending upon it certain histological alterations, go on at temperatures far below the freezing point. They have proven, also, that even four years at such low temperatures are not sufficient to render the flesh of chickens germ-free, and, judging from the work which has already been reported by various investi- gators, it seems probable that both bacteria and enzymes will resist intense cold for long periods, only thawing and a rise in temperature being needed to cause a rapid resumption of activities. Some recent contributions to our knowledge of bacterial life at tem- peratures below zero would indicate that, if the organisms are pro- @Loc. cit. 6 Uber den Einfluss niedriger Temperaturen auf die Funktionen des Magens, Zts. Biol., 18 (Neue Folge, 10): 433. ¢ Verhandlungen der Wiirzburger physiol. med. Gesell., 1872, N. F. 2: 122. d Pfliger’s Archiv gesam. Physiol., 14: 395. € Versuche zur vergleichenden Physiologie der Verdauung mit besonderer Beriick- sichtigung der Verhaltnisse bei den Fischen, Untersuch. aus dem physiol. Inst. zu Heidelberg, 2: 395. f Uber die Magen- und Darmverdauung bei einigen Fischen, Inaug. Diss. Kénigs- berg, 1878. g Amer. Chem. J., 1900, 24: 491. 99 PRELIMINARY COLD STORAGE STUDIES. vided with their natural environment, they will multiply when the medium is in a frozen condition. The experimental work which has been done in the past has used chiefly the usual laboratory media and pure cultures of laboratory-grown bacteria. It is perfectly possible that such conditions, since they are not comparable with those natu- rally prevailing, will lead to some erroneous results, and in view of recently demonstrated facts it becomes necessary to attack the prob- lem of bacterial development in flesh foods from this point of view before the assumption can be accepted that temperatures below freez- ing guarantee a freedom from bacterial activity. Observations on the growth of bacteria under the conditions of the very low temperature cold-storage houses, such as almost universally prevail in the United States, are entirely lacking. What informa- tion is to be had on the growth of bacteria in flesh when cold-stored comes from abroad, chiefly from Germany, where the temperature of the ‘‘Kiihlhaus’’ rarely reaches 0° C. and is commonly several degrees above it. Exposed to such temperatures there is a unanimity of opinion regarding the “‘ripening”’ of the flesh, and the tenderness and flavor acquired in the course of it. To what this*maturation is due, however, is not so well settled. Glage * would ascribe much of the flavor to ‘‘aroma-producing”’ bacteria which develop best at low temperatures; Miiller,® on the other hand, believes that the process is essentially dependent upon the enzymes of the flesh itself. He holds that the temperature of the chilling room prevents putrefac- tion, and, therefore, all those poisonous properties dependent upon putrefaction, while it assists natural autolysis. Neither does he con- sider ° that the changes in the ripening of meat are the early stages of putrefaction. The observations made under commercial conditions he has rein- forced by a study of freshly killed, bichlorid-washed fish, which was kept at 0° C. After 5 days there was an unpleasant taste and a characteristic odor, both of which appeared in 2 days when kept at 12° C. Schmidt-Nielsens kept a carp packed in ice for 14 days, and, though bacteria-free, it had, at the expiration of the above period, so unpleasant a taste that it was unfit for food. Miller determined the amount of nitrogen soluble in water in both mammalian and fish muscle kept at 0° C., concluding therefrom that an autolysis proceeds. He found that muscle loses its elasticity, becomes tender, and the clear red color changes to an opaque, dark red. The odor after 3 days at 25° C. or at 0° C. after 14 days is strongly acid. a@Zts. Fleisch- Milchhygiene, 1900-1901, p. 131. © Der Reifungsprozess des Fleisches, Zts. Fleisch- Milchhygiene, 1904, 14: 217 and 337. ¢ Archiv Hyg., 1903, 47: 127. MARKET COLD-STORAGE CHICKENS. 93 In 1897 Gautier® published the account of a chemical and _his- tological examination of frozen flesh, chiefly of that sent to France in a frozen condition from South America and the United States. He stated that the flavor of the frozen muscle was never quite as good as the fresh. That the juice when exuded was always more abundant than from the fresh and that it contained globulins, albu- mins, peptones, and organic and inorganic constitutents. Among the most recent chemical studies of such flesh is that made by Rideal,? who determines the ratio of nitrogen to the total solids in both chilled and fresh meats and, finding them the same, excludes decom- position. Artificial digestion gave identical results with frozen and fresh muscle. He states that there were no signs of incipient decom- position. Neither bacteriological nor histological examinations were reported. However, this statement is made: ‘‘The tenderness of meat which has been frozen has been attributed to the slow action of sarcolactic acid, and the Hoses of the intermuscular tissue promotes rapid fagchapaeen. © Martel,° in a recent article on the cold storage of foodstuffs, favors cold rooms rather than a temperature sufficiently low to freeze, believing the latter only necessary when transportation for long distances is to follow. He states that bacteria do not readily pene- trate the muscle, about 10 days being required to carry them within 1 cm of the surface, but that there is a marked autodigestion of the cell contents, due to the action of the cell ferments, which proceeds easily at 2° to 3° C., though below zero such action is stopped. As to the quality of the meat after storage he believes it to have an improved flavor if kept at 2° to 3° C. and not allowed to freeze. The cause of this improved flavor is ascribed to the aroma bacteria, which he considers desirable. Accompanying the change in taste there are, according to Martel, alterations in the muscle fiber which consist in a change from trans- lucent to opaque and from brilliant to dull, as well as from tough to tender. The reaction of the fresh muscle is neutral, but becomes acid as coagulation proceeds. In cold storage at 2° to 3° C. about 8 hours are required for a strong acidity to develop, and an odor, aromatic but not at all putrefactive, makes its appearance at the same time. When muscle is fresh it is very difficult to extract any fluid. After 3 days in storage flesh yields much juice. Micro- scopic examination shows an annihilation of the striations and abundant granulations. a Les viandes fraiches et congelées. b Cold Storage, 1907. ¢ Conservation et maturation des viandes emploi du froid industriel, L’ Hygiene de la viande et du lait, Vol. 1, Nos. 1 and 2, 94 _ PRELIMINARY COLD STORAGE STUDIES. Discussion OF RESULTS OBTAINED IN THE BUREAU OF CHEMISTRY. HISTO-CHEMICAL CHANGES IN COLD-STORED CHICKENS. Gautier * states that the muscle fibers of cold-stored flesh remain unaltered except, perhaps, for a slight pulling apart of the indi- viduals. The marked and deep-seated changes, the occurrence of which has been demonstrated in connection with the study in this laboratory, of chickens preserved by cold are, therefore, of interest, and it seems probable that the solving of the problem of the cause, sequence, and ultimate result of such changes would throw much light on the whole question of the various alterations undergone by flesh at low temperatures. While the various dyestuffs which have been used to differentiate the tissues preparatory to microscopic study have not, as yet, served as exact microchemical reagents in the sense of classification of compounds, they have very positively indicated an alteration in the chemical character of certain mor- phologically distinct elements. The characteristic green of the normal, fresh muscle fiber, when stained as previously described, is very decidedly altered by long keeping at low temperatures, so much so that finally it is almost entirely replaced by dirty yellow and brown greens, or even by orange tints. The white fibrous connective tissue, staining a brilliant blue in the fresh chicken, in the cold-stored takes on a greenish tint. The material which exudes from the muscles after keeping for even a comparatively short time has, at first, almost the same staining reaction as the fiber itself, but it changes gradually, until it is a dirty brownish or bluish green. The irregular staining of the fiber would also indicate the presence of a progressive chemical alteration. The rupture of the sarcolemma and the extrusion of the muscle substance are not merely mechanical alterations due to freezing and thaw- ing. If such were the case the progressive effects which are so clearly traced in fowls kept for periods of varying lengths of time would be wanting. The differences between the chicken muscle frozen for 48 hours and that of the fresh bird lies chiefly in the size of the spaces between the fibers or bundles of fibers, and deep-seated changes are not seen. The histological alterations which have been traced are confirmed by the differences noted between stored and fresh chickens in the distribution of the protein nitrogen. It would seem probable that for the chickens stored 2 years and 4 years, respectively, the greater part of the change is due to enzyme action, not only because the tissue shows but few bacteria either living or dead, nor because the histological degeneration proceeds differently from that observed when ordinary temperatures prevail, but also because of the changes @ Loc. cit. ; MARKET COLD-STORAGE CHICKENS. 95 in the relative distribution of the protein nitrogen. The fowls in stor- age 14 months give indications of more marked changes, chemically, than did the others examined, considering the length of the storage period, but such changes may be due to the number of organisms which were found in a living condition in the tissues and which may have been present when the fowl was stored. Because these chickens have an unknown history preceding their entrance into the cold-storage warehouse it is impossible to say whether the numbers found represented an increase or a decrease during the storage period. Such questions can only be answered by the study of chickens of known and of a strictly comparable history. Neither can this report deal comprehensively with the question of the migration of organisms from the intestine into the edible portions. Judging from the ravages undergone by the walls of the gut during storage it could offer but a slight barrier to active bacteria. - From an investigation which is now in progress to determine the resist- ance of intestinal organisms to cold when in their natural environ- ment it appears that they do remain alive in large numbers. They are, however, of the varieties which develop best at 20° C. instead of body heat, agreeing in this with the behavior of the naturally occur- ring milk organisms when kept under like cireumstances.? The fact that the organisms which have been found are not gas pro- ducers in dextrose media would argue against possible migration. The location of the bacteria in the tissues, on the other hand, would indicate its possibility since there are fewer in the muscles of the breast than in the inner thigh muscles, which in the chicken le closely adherent to the body wall. CHANGES IN FAT. The decomposition of the fat of the chicken is much more pro- nounced than is the decomposition of the protein, and in the tracing of the changes which occur in this tissue not only bacteria and enzymes must be taken into account, but light and air must also have due consideration. It has been held that the splitting of fat into acid and glycerol is the cause of rancidity. More recent studies would indicate that, while such a splitting ordinarily accompanies the condition known as “rancid,’’ it is not the real cause of it, but that rancidity is due to _ the action of air and light on fats which have been previously split by enzymes acting in the presence of moisture.? The part played by bacteria in the decomposition of fats in their natural environment is not a minor one, though the réle to be assigned to them regarding «Pennington, loc. cit. 6 Lewkowitsch, Chemical Technology and Analysis of Oils, Fats, and Waxes, 3d ed., 1904, p. 22, 96 PRELIMINARY COLD STORAGE STUDIES. the production of rancidity is still a mooted question. Since bacteria will not propagate in pure, dry fat, but must have present suitable nitrogenous foodstuffs as well as moisture, the conditions for their activity, as pointed out by Lewkowitsch, are the same as for the activity of enzymes, hence it is exceedingly difficult to say where the action of either the one or the other ceases, but it would seem proba- ble that fat splitting can be ascribed to both enzymes and bacteria, whereas the decomposition of the free fatty acids or of the glycerol must be referred to bacteria alone. The researches of Kastle and Loevenhart on lipase ¢ have established the wide distribution of the enzyme in nature and its stability as well as its adaptability to its environment. It was kept dry and moist, and at room temperatures and in cold storage, retaining its activity for months. It hydrolyses most rapidly at 40° C., at which tempera- ture 11.29 per cent of fat splitting was noted; at 10° C. the amount was 3.89 per cent; at 0° C., 2.26 per cent, and at —10° C., 0.70 per cent, a quantity which, given weeks and months in which to aug- ment, might easily reach notable proportions. The enzyme lipase shows a decided preference for fats of higher molecular weight, wherein it differs from the usual acids used to induce hydrolysis. It is also able, under favorable conditions, to reverse its action, and syn- thesizes ethyl butyrate, for instance, from alcohol and butyric acid. Hanriot® states that lipase can form monobutyrin from butyric acid and glycerin and the higher the molecular weight of the acid the more easily is the synthesis accomplished. Such facts indicate a most important and far-reaching physiological réle for the enzyme, as is discussed at length by Loevenhart, who finds it in considerable quantities wherever fat synthesis is taking place in the living tissue, as in the subcutaneous fat and in the secreting mammary gland. Between the enzyme studied by Loevenhart, in the milk gland, and that occurring in milk, Gillet? finds a most important difference, namely, that the latter splits monobutyrin but not ordinary fats. In accord with such a train of thought comes the work of Connstein, Hoyer, and Wartenberg,’ who found a great variation in the ability of lipase to split fats from different sources. Butter is very resistant, owing, probably, to its high percentage of acids of low molecular weight, since triacetin gave 0.4 per cent, tributyrin 9.5 per cent, and ~ triolein 50.6 per cent of free fatty acid when all were treated under like conditions. It is also noteworthy that the reaction is much more «Concerning Lipase, the Fat Splitting Enzyme, and the Reversibility of its Action, Amer. Chem. J., 1900, 24: 491. 5 Sur la reversibilité des actions diastatiques, Comptes rend. soc. biol., 1901, p. 70. ¢ Amer. J. Physiol., 1901-2, 6: 351. d Existe-t-il une lipase dans le lait?—J. physiol. pathol. general, 1903. éThe Enzymic Decomposition of Fat, Abs. J. Soc, Chem. Ind., 1902, 21: 1541, MARKET COLD-STORAGE CHICKENS. 97 rapid in the beginning of the experiment than when there has been an accumulation of the products of activity. Such wide and varied abilities appertaining to an enzyme as are indicated by the foregoing summary demand that account be taken of its action, wherever natural changes of fat are in progress, and its ability to retain its activity for long periods of time and to work at low temperatures makes its réle in fat changes m cold storage an important one. Since enzymes of varied abilities have been traced to a bacterial origin it is quite reasonable to infer that those having some action on fats would be so produced. Schreiber,“ who has studied earth bacteria, was unable to find ferments, and considers the decompo- sition of fats to be dependent upon vital functions. It is then affected by all the conditions which affect life. Schreiber states that anaerobes split fats slightly, but their activity ceases at that point and is feeble at best. Rahn® denies that anaerobes have any action on fats, but admits that they can decompose glycerin. Both of these authors report work on certain molds—Mucor, Penicillium— and find that they cause both a rapid and a complete decomposition. Among bacteria B. fluorescens liquefaciens shows a fat-decomposing power closely following that shown by molds. Rahn° has deter- mined this activity as expressed by the iodin number, which was considerably increased for palm fat, stearin, and butter fat. Laxa? has recorded analyses covering the values usually determined in fat examinations using butter fat with molds, yeasts, and fat split- ting bacteria proper. The latter, with the exception of B. fluorescens liquefaciens, commonly cause a decrease in the iodin number; the other fungi noted usually increased it. The Reichert-Meissl number was but little changed. The acid value increased for both molds and bacteria. The saponification number decreased with both. The index of refraction in many instances fell when the iodin number rose, and when the latter decreased the refractive index remained stationary. The Hehner number for molds showed a rise. It will be observed that there is a distinct difference between the action of molds and of bacteria on fats. This is again expressed in the work of Girard, who believes that Aspergillus and Penicillium act by way of an enzyme and Laxa’has demonstrated by experi- ment that such is the case. With substances as complex as are fats in their natural environment and with forces as delicate as fungi @¥ettzersetzung durch Microorganismen, Arch. Hyg., 1902, 41: 328. bDie Zersetzung der Fette, Centrbl. Bakt., 2 Abt., 75: 53. cMOee Clb. to 422, dUeber die Spaltung des Butterfettes durch Microorganismen, Arch. Hyg., 1902, 41: 118. € Loc. cit. 49078—Bull. 115—08——7 - 98 PRELIMINARY COLD STORAGE STUDIES. and enzymes, and with such universal factors as. air and light to be considered, it is not to be wondered at that the whole question of fat decomposition in nature is still greatly in need of elucidation. The literature on the subject, as indicated in the foregoing discussion, is chiefly concerned with plant oils and butter fat and with fats freed from the tissues normally occurring with them. Almost invariably, also, the studies made have been conducted at body or at room temperature. The discussion of the changes in the fat of cold- stored chickens as indicated by the variation between their values and those of the fresh birds, which values are given in another section of this report, becomes therefore a difficult task. Light, as a cause of decomposition, need scarcely be considered; air, at least for the superficial fats, is probably a factor and would tend to increase the acidity. So great are the differences between the cold- stored and fresh chickens when the acid value, saponification number, and ester number are considered that probably not only air but en- zymes as well, and perhaps bacteria too, have all played a part in the alteration. The lowering of the iodin number rather argues for a certain amount of bacterial action, as does also the decrease of the saponification number, since the available contributions on the subject agree in assigning such results to bacteria. The change in the Hehner number due to bacterial action is not discussed to any extent in the literature. Laxa,¢ in his study of butter, makes two determinations, one for Oidiwm lactis and the other for B. fluorescens liquefaciens, finding an increase in both cases. The Hehner number for fresh chick- en fat is lower than is usually found for animal fats, and after cold storage a further decrease is observed. It is difficult to explain the cause of a rise in the Hehner number as a result of fungoid action, since that is essentially katabolic, and not at all of the character to produce acids of a higher type than those normally present, whereas - the decomposition of insoluble acids with the production of soluble forms is perfectly logical and quite in line with the observations of Rahn and Krueger. SUMMARY OF RESULTS. (1) Macroscopic observations of fresh and cold-stored chickens show that certain plainly visible differences exist between the two classes, which differences are progressive, depending on the length of the storage period. (2) Chemical analyses of fresh and cold-stered chickens, wherein were determined the total nitrogen for both dark and light meat and its distribution between the classes of compounds commonly accepted as the result of protein hydrolysis, the various values from which a knowledge of the composition of fat may be obtained and such @Loc. cit. GENERAL REVIEW OF THE INVESTIGATION. 99 minor constituents as water, ash, etc., have served to show that for the protein distribution there is a slight variation in the cold-stored product from the fresh, and for the fat values a wide variation. (3) A histological examination of the muscle of both fresh and cold-stored chickens shows a marked and a progressive change in the structure of the fibers which is deep-seated and after long periods renders the tissue almost unrecognizable. Selective microchemical differentiation of the tissues confirms the chemical change found by eross analytical methods. (4) A bacteriological examination of fresh and cold-stored chickens reveals the presence of appreciable numbers of organisms, calculated on the gram basis, in the edible portions of those preserved by cold, though the numbers were not large. In fresh fowls the same tech- nique gave no bacterial growth. GENERAL REVIEW OF THE INVESTIGATION. In reviewing the details of this preliminary work it must be remem- bered that some of the inferences drawn may be modified. by future investigations, although the fact that certain changes take place in the food materials examined after definite periods of storage is well established both by the organoleptic and by the chemico-bacterio- logical tests and is further confirmed, in the case of the chickens examined, by the histological studies. The principal lines of investigation reported, namely, on eggs, quail, and chickens, conducted at Washington under known conditions of storage, and the more complete study of chickens stored under market conditions at Philadelphia, are summarized in the following pages, brief reference also being made to the conclusions drawn from the milk investigation previously reported by Pennington. ORGANOLEPTIC TESTS ON CHICKENS AND QUAIL. The general results of the organoleptic tests of the stored fowls and birds leads to the conclusion, first, that even after three months, before cooking, it is not difficult to distinguish, by the appearance, color, and odor, a freshly killed bird from the one that has been in cold storage. The shriveled condition of the eye and the skin and the generally dilapidated appearance of the bird are very significant and distinct. After cooking, however, within a period of three months, there is much disagreement on the part of the jurors respect- ing the proper classification thereof. This is especially true as to distinguishing between the drawn and the undrawn birds, where the ' variations in judgment are of such a character as to lead to the belief that it is impossible within that time, from the taste, odor, and smell of the cooked bird, to determine which one had been drawn and which - was undrawn at the time of storage. The possibilities of determining 100 PRELIMINARY COLD STORAGE STUDIES. which is the fresh bird are very much greater, even after a lapse of three months, although occasional mistakes may be made in this respect also. At later dates, such as at the end of six months, nine months, or twelve months, the difference between the birds becomes more and more pronounced, so that it may with certainty be said that even after cooking one would rarely confound a fresh bird with one which had been in storage six months. At that date the flavor and the general character of the meat have so deteriorated that it is not difficult to distinguish between the fresh and the stored fowl. Even at this date, however, there is some difficulty in distinguishing between the drawn and the undrawn bird. At the end of a year or more it would be quite impossible to make a mistake in most cases between the fresh and the stored birds. | Summing up the organoleptic properties, it may be said that for a short time, possibly six weeks or even longer, there is no perceptible change produced in a chicken by having it frozen. There certainly does not seem to be any evidence that it is better, and there is no convincing evidence that it is any worse. After three months, how- ever, the fresh chicken is easily distinguished by its properties, as a rule, from the cold storage chicken, even after cooking, and to an absolute certainty before cooking. This distinction between the fresh and the stored bird becomes more and more marked as the time of storage is increased. In so far as the drawn and undrawn chickens are concerned there is much less certainty of being able to distinguish between them. However, 70 per cent of the jurors were able to pick out the undrawn bird by its stronger odor and taste after a storage period of from six to fifteen months, but at the test representing 18.5 months’ storage the two birds were about equally dry and tasteless. The general conclusion is, therefore, that in the case of frozen birds there is no indication of any improvement in quality, that is, in taste, odor, or flavor, during cold storage. There is a deterioration which is noticeable, even at the end of three months, and becomes more marked as the time of storage grows longer. Hence, without any ref- erence whatever to the question of wholesomeness, cold storage pro- longed for six months or more appears to be distinctly detrimental as far as taste, flavor, and palatability are concerned. BACTERIOLOGICAL AND CHEMICAL INVESTIGATIONS. MILK. As a part of this investigation certain studies have been made on the changes taking place in milk when kept at low temperatures. The detailed report of this study has already been published,” hence aJ. Bioi. Chem., 1908, 4: 353, Bacterial Growth and Chemical Changes in Milk Kept at Low Temperatures, M. E. Pennington. GENERAL REVIEW OF THE INVESTIGATION. Oe there will be included here only the conclusions reached, which are as follows: SUMMARY OF RESULTS. Bacteria in milk increase in numbers when the temperature is maintained at or a little below 0° C. This temperature is below that ordinarily assigned as the limit of bacterial multiplication. Milk has been kept for periods ranging from a few days to almost two years at a temperature of 29° to 31° F., and also at 32° F. It has been kept in a packages of ten quarts and one quart. It has been the cleanest milk obtainable, by the most carefully enforced refinements of modern dairying; and it has also been market milk produced in the ordinary dirty stable and subjected in transit to the usual careless handling. Bacterial growth at the end of a week, even in the cleanest milk, which contained as low as 300 organisms to the cubic centimeter, was pronounced. There was a steady increase in the number of organisms for five or six weeks, and at their maximum they numbered hundreds of millions. Occasionally they passed the billion mark per cubic centimeter. Continued exposure to a temperature of 29° to 31° F. causes, after a lapse of from seven to twenty-one days, the formation of small ice crystals which gradually increase until the milk is filled with them and there may be an adherent layer on the walls of the vessels. The milk does not freeze solidly. In spite of the fact that the milk was a semisolid mass of ice crystals, the enormous increase in bacteria which this study shows took place. Though the bacterial content was numerically in the hundreds of millions per cubic centimeter there was neither odor nor taste to indicate that such was the case. Neither did the milk curd even on heating, and it was not until the bacterial content began to fall, and organisms of putrefaction were under way, that the use of the milk for household purposes would, to the ordinary observer, become contra-indicated. ; A classification on a chemical basis of the organisms occurring at these low tem- peratures shows that there were constantly present bacteria which formed acid and bacteria which acted upon proteid. There were also neutral organisms, which formed neither acid nor alkali and did not act upon gelatin. The acid-forming organisms were generally in relatively smaller numbers than are found when milk is kept at higher temperatures, and the liquefying organisms were more numerous. Certain spe- cies, such as B. formosus, B. solitarius, and B. raveneli, were especially resistant to cold and frequently were the predominating species, or almost in pure culture at the close of the experiment. A very marked difference in both the number and kind of organisms which devel- oped on the plates was noticed, depending upon the temperature at which the plate was incubated. In certain experiments the maximum number grew at 37° ©. In others the temperature at which the milk was stored served best for colony formation. The relative number of organisms growing at 37° C., 20° C., and 0° C., ora little below, varied greatly also with the length of time that the milk had been kept in storage, the organisms developing at body temperature being ordinarily greatly in excess at the beginning of the experiment and diminishing until near its close, when a sharp rise was apt to take place. The determination of the acidity showed that after a few weeks a much higher acid content was reached than is ordinarily required for the spontaneous separation of curd, which, however, seldom happened. Milk having this high acidity, when placed in an ordinary ice chest, increased in acid content but did not curd for days alter exposure to the higher temperature. The chemical study of the proteid of milk in cold storage showed that the casein was rapidly digested until finally more than 50 per cent of it was changed to soluble compounds. Caseoses, amido acids, and probably peptones, increase apparently at 102 PRELIMINARY COLD STORAGE STUDIES. the expense of the digested casein. The rapidity with which this digestion takes place varies in different samples, but at the expiration of two weeks it is pronounced. What the effect of the low temperatures is on the carbohydrate constituents of the milk remains for further study. That an interesting decomposition, and one which varies from that occurring at higher temperatures, takes place is indicated by the very high acid content of the milk noted. throughout this investigation. Similar studies, conducted on samples of very fresh milk kept at the temperature of the laboratory (about 18° to 22° C.), show a very decided difference chemically from the decomposition of milk in cold storage. Bacterial growth at room tempera- ture is, of course, rapid and profuse. The acid-forming organisms are, as has been found by other observers, in high proportion and the liquefying organisms are rela- tively lower. The chemical change observed is, by comparison with that occurring in cold storage, almost nothing. At the curding point only about 1 per cent of the casein has been changed to soluble products, and spontaneous curding is observed, ordinarily, when the acid content falls between 23 and 28 cc of tenth-normal sodium hydrate per 100 cc. CHICKENS. The results obtained show unmistakable evidences of bacterial activity during the period of storage. While naturally the frozen eondition of the flesh prevents to a great extent bacterial migra- tion, it does not prevent absolutely bacterial growth. It is perfectly certain from these experiments, as well as from others, to which ref- erence has been made, that a degree of cold, equivalent to that sus- tained in ordinary cold storage, does not by any means destroy bac- teria. The effect is simply to produce a state which may be described somewhat broadly as suspended animation. It is also certain in some instances that the number of bacteria may be diminished during cold storage, showing that while cold does not kill all species of bacteria it may hold certain forms in suspended animation so long that they are finally killed. Attention should be called also to the fact that as this line of work was entirely new, a great deal of the time has been devoted to deter- mining the best methods for the investigation and establishing the general principles on which it should be based. It is evident, there- fore, that the deductions which are to be drawn here are only to be considered of a tentative nature and should not be regarded as the final word on the subject. Too few specimens have been examined to make the data sufficiently extensive to be relied upon as a final basis of conclusions. The experiments have undoubtedly been car- ried on under conditions which are more favorable than those exist- ing in the markets in general. The fowls on which the work conducted at Washington was done were specially selected, killed with the greatest care, packed with the utmost precautions, and in every way subjected to the cleanest con- ditions. On the contrary the fowls which are ordinarily placed in cold storage are not selected fowls, they have not been killed and prepared with special care, nor are any special means taken to pre- GENERAL REVIEW. OF THE INVESTIGATION. 108 vent possible infection. It is evident, therefore, that the data which have been obtained from the bacteriological study of the market cold-stored chickens, as conducted at Philadelphia, must be accorded full value, and the acknowledgment made that with ordinary cold storage of fowls under market conditions the bacterial flora is hkely to be very greatly increased. In the examination of the market cold-storage chickens there was observed a very definite correlation between those fowls in which the quantitative bacteriological findings indicated a relatively large number of organisms and the gross chemical analysis of the bird so far as the degradation of proteids was concerned. Here also the bac- teriological and chemical findings were abundantly confirmed by his- tological sections of the tissues, which showed an invasion of the individual fibers by masses of organisms which had undoubtedly altered the chemical composition of the muscle fibers themselves to such an extent that finally their chemical integrity was absolutely lost. For the detailed summary of the various phases of the inves- tigation of market cold-stored chickens reference should be made to the close of the special report on this subject, page 94. Among other considerations the proper removal of the viscera from the cavity of the drawn fowls is a matter of great consequence in case they are to be kept in this condition. In point of fact, however, a very small percentage of the fowls which are placed in cold storage are drawn, so that the precautions for the removal of the intestines without danger of infection are of no very great consequence. In drawn fowls, however, such as were half of those stored for the Wash- ington experiments, the utmost care was exercised to prevent infec- tion from the intestinal contents durimg drawing. This is a danger which the advocates of drawing fowls have, as a rule, overlooked. It is evident that without proper precaution very grave danger of intes- tinal infection may attend the drawing of fowls. Attention must also be called to the fact, which seems to be well established by the investigations, that food products which have been kept a long while in cold storage, even if they are sterile, are much more sensitive to infection and the rapid development of dele-. terious substances due thereto than when in the fresh state. It is com- monly conceded to be a general practice in withdrawing cold-storage chickens or other birds from the warehouse to place them in very cold water until the flesh is thawed. This has a double purpose of preventing too rapid thawing, which would be somewhat destructive to the tissues, and at the same time of permitting the absorption of the water which has been lost during the time of storage. During this period it is evident that the carcasses must be subjected to infection. Subsequent to this preliminary treatment the chickens are placed upon the shelves for sale and may remain there a long time, depending 104 PRELIMINARY COLD STORAGE STUDIES. upon the state of the market, before finally reaching the consumers’ hands. After this they remain for some hours in the hands of the consumer before cooking. It is easy to conceive of a period of three or four days covering all of these various operations. All this time these materials, so sensitive to infection, are exposed in various ways to danger. . In the consideration of the bacterial studies it must be borne in mind that it is not impossible to find bacteria in the tissues of the fresh fowls. While it is commonly accepted that the general circu- lation of animals in a state of perfect health is free from bacteria, it is quite certain that in abnormal conditions bacteria may exist in the organs of the body and even in the blood, hence the detection of bacteria in the tissues of a cold-storage fowl, or any of its organs, may not be conclusive proof that these bacteria were developed during cold storage, since they may have been there in the original state in which they were packed. The findings of the macroscopic investigations emphasize and con- firm the organoleptic tests previously discussed in that there was observed a decided fecal and irritating odor, suggesting rancid fat, in the case of the undrawn fowls long in storage, existing coincident with the taste which enabled the jury to distinguish between the two methods of dressing. On the other hand, in the drawn fowls there was a tendency to a rancidity of the fatty portions of the abdominal region and a development of an odor somewhat similar to that pro- duced by a butyric fermentation. In general, however, in so far as the bacterial investigations have proceeded, there is little choice between the drawn and the undrawn fowl. It is possible that on longer keeping there will be developed, in from one to three years, im- portant differences between the drawn and undrawn fowls which will enable them to be distinguished, both by their bacterial flora and by their organoleptic and chemical properties. HISTOLOGICAL STUDIES. EGGS. _ As is well known, the flavor of eggs begins to deteriorate very soon, even when they are kept cold. In the storage of eggs, moreover, the temperature must be kept above the freezing point—that is, the freezing point of an egg. The egg, on account of its constitution, does not congeal at the temperature at which water becomes ice. It is possible to cool eggs considerably below 32° F., at least for a short time, without freezing them. Nevertheless, it is not safe, as a rule, to go very much, if any, below the freezing point of water. Kept in this way, eggs undergo certain changes, not only of a histological nature, but of a chemical character, which are now under investigation. Certain constituents of the egg have a tendency on storage of this GENERAL REVIEW OF THE INVESTIGATION. 105 kind to become crystalline. In the course of a few months eges which are kept in cold storage are found to develop small rosette crystals in the yolks. These, it has been found, occur most frequently in the outer portion of the yolk. ‘The size of the crystals is found to be from 18 to 109 in diameter, and the form is shown in Plate III. The exact character and composition of these crystals has not yet been ascertained, as they constitute a relatively small percentage of the entire mass, and hence their isolation in sufficient quantities for analysis is attended with extreme difficulty. Many attempts, as has been seen from the detailed statement in the preceding section of this report, have been made to isolate and prepare these crystals for identification. It is perhaps possible that they belong to the class of substituted fatty bodies, but no definite statement can be made except that it seems that they are not tyros. The observa- tions of these bodies seems to be entirely new, as no account has been found of them in other publications. It is probable, therefore, that their existence may be regarded as one of the means of distin- guishing eges which have been a considerable time in cold storage from fresh eggs. CHICKENS. Quite as important, and perhaps even more instructive than the studies relating to bacterial growth at low temperatures, are the studies of the changes which take place in the tissues of the material, and which can be recognized under the microscope. Changes in tissue are usually due either to fermentation or to enzymic action, if the two terms are to be used in any different sense. It is generally conceded by investigators at the present time that the actual changes in the structure of the tissues are not so much due to direct action of the bacteria as to the enzymic ferments which bacteria secrete or produce or which exist in the normal cells of the tissue itself. Hence the action of enzymes may be regarded as the last step of fermen- tation. The studies in histological changes were undertaken on market chickens which had been stored for varying lengths of time at about 13° F. One difficulty in connection with a general investigation of this kind is that if chickens are selected at random, or purchased in the open market after having been dressed, a good deal of uncer- tainty must exist concerning their previous history and the condi- tions to which they have been subjected. Commercially it is well known that the interval which exists between the period of slaughter and the period of storage is of varying length. The temperature and other conditions of environment to which the slaughtered chickens are subjected in this interval are far from uniform. The method of transportation may have much to do with the character and extent of the changes before cold storage in a warehouse begins. 106 PRELIMINARY COLD STORAGE STUDIES. Chickens may be, for instance, prepared for shipment at distant points and sent in refrigerator cars either frozen or at a low tem- perature on a voyage requiring a long period of time before they finally reach the depot in which they are preserved until a demand for consumption arises. It was necessary, therefore, in all studies of market chickens to begin the work with fowls whose history was known so that they could be followed, not only from the time of slaughter, but even before that time. The different breeds to which the chickens belong, the methods of hatching and raising, the charac- ter-of the feed which they eat—all may have important bearings upon histological and chemical, as well as upon bacteriological, data. Hence the desirability of the practice which was finally adopted, not only of studying chickens in a commercial way, but also of having those of known breed and history. Manifestly it was impracticable to study microscopically all of the edible parts of the bird at this time, hence it was decided to select for comparative work the fibers of the large muscle of the breast, the Pectoralis major, which is usually considered the choicest food portion, and which is apparently the part which keeps longest and best. In order that the character of the changes may be more readily appreciated, they have been reproduced by microphoto- graphs and by colored plates, so that the reader who is not skilled in microscopic work nor in histological investigations may be able to see at a glance the character of the changes which have taken place. (See Pls. VI to XI.) The sinuous outline of the fresh fibers soon dis- appears in cold storage, and the fibers are not by any means so flex- ible as they were in the fresh state. The most obvious change, however, which takes place during this interval is manifested as a structureless, granular substance which hes between the individual fibers and between their aggregations which are known as bundles. The origin of the substance is in the fiber itself, and in the earlier periods of storage its composition, according to reactions obtained with various selective dyestufis which afford exceilent microchem- ical reagents, is not very different from that of the normal fiber. As the storage period is lengthened, however, most marked differ- ences between the normal and cold-stored tissues, as exhibited in the staining reactions, make their appearance, and such changes, whether referable directly to bacterial or enzymic action, are essentially chemical, and deal with the fundamental principles composing flesh foods. The results obtained from this microscopic study of the his- tology of market cold-stored fowls would seem to point the way to a series of investigations, which has already been begun and from which it is believed will accrue a knowledge of the obscure chemical changes undergone by protein, to elucidate which the present gross chemical methods are entirely inadequate. - = ; : 4 GHNERAL REVIEW OF THE INVESTIGATION. 107 Such changes as were observed in the flesh of these cold-stored fowls make it especially desirable that there be carried out certain pharmacological experiments, using preparations of cold-stored tis- sues. It is held by some of our most eminent medical authorities that many severe intestinal disturbances are traceable to cold-storage animal products, particularly when the viscera are not removed before storing. Hemmeter®@ says in this connection: ‘‘I have per- sonally observed numerous cases of sudden and severe auto-intoxica- tion from the gastro-intestinal tract which I could interpret in no other way but that they were due to the ingestion of cold-storage food.” In view of the indisputable fact that changes do take place in flesh at low temperatures, and considering the authority lent to the above statement by the prominence of its author, it would seem most necessary that further experiments based on strictly scientific principles be prosecuted looking toward either its refutation or expla- nation in order that the evil may be remedied, if it exists. The intestines, which are left in situ in storage birds, show a very marked degeneration. Their muscular walls grow thinner in cold storage until they are the merest remnants, which threaten to disappear alto- gether and which even very careful handling may easily rupture. This degeneration is noticeably active in the muscular rather than in the cellular tissues of the intestines. This is important when it is considered that the bacterial flora of the intestinal contents will, of course, contain any pathogenic germs which usually accompany the colon bacillus.. Hence the perforation of the walls of the intestines, which apparently takes place by continued digestive processes even in cold storage, would open the way for a rapid migration of such bacteria on thawing and previous to cooking. Thus it is quite pos- sible that dangerous bacterial organisms might be translated to the edible portions of the fowl through the perforations of the intestines in the period between thawing and cooking. This degeneration of the walls of the intestines must, therefore, be regarded as highly significant. a@ Memorial number of the Maryland Medical Journal, June, 1905, 48: No. 6. APPENDIX. TREND OF LEGISLATION REGULATING THE COLD STORAGE OF FOODS, AND OPINIONS EVOKED THEREBY, ESPECIALLY WITH REFERENCE TO DRAWN AND UNDRAWN POULTRY, ETC. GENERAL DISCUSSION AROUSED BY PROPOSED CHICAGO ORDINANCE. In 1906 an ordinance was introduced in the Chicago council to exclude from the city trade all undrawn poultry or animals as food products. In the course of the discussion which this bill evoked Runnels and Burry, attorneys for the cold-storage warehouse interests, issued a pamphlet dealing not only with the question of drawn and undrawn poultry, but with cold-storage conditions in general, and from which the following extracts are taken. In regard to the quantity of material stored they state: About twenty million pounds of poultry are cold stored in Chicago each year, and there are stored proportionate amounts of butter, cheese, meats, game, eggs, and other food products. In a letter from a large western firm the statement is made: We bring to the city of Chicago for storage annually 6,000,000 pounds of poultry, 300 carloads of eggs, and 100 carloads of butter, * * ™* 5 per cent is sold for Chi- cago consumption. The other 95 per cent is sold for consumption in cities outside of the State of Illinois or abroad. During the month of January of the year 1906 this firm shipped to Liverpool and London 1,000,000 pounds of poultry. The business of this firm during the year then just past (1905) had exceeded $1,500,000. In dealing with the question of the length of time the various cold- stored products should be or may be carried, the pamphlet cited makes the following statements: Eggs and apples, which are perhaps stored more extensively than any other com-_ modities, can not by any system of cooling or refrigeration be kept a year in cold storage, and they are usually taken from storage in a much shorter time than that. Probably few apples are kept over six months in cold storage. * * * Hggs are put in storage during the months of April, May, June, July, and a few in August. April and May eggs keep best. In selling them the owner sells the latest eggs first, so that July and August eggs are never in storage more than from two to four months, while April and May eggs are closed out usually during December and January. Some eggs remain, therefore, in storage more than six months, but the rule is that they are to be disposed of by the first of February. - 108 APPENDIX. 109 Poultry is sent to market, dressed and stored, during the months of October, Novem- ber, and December. The amount so stored, killed when poultry is in its best con- dition, is intended to supply the market with the best goods obtainable until the new stock comes in during the next receiving season. Necessarily, therefore, part of this stock is carried for more than six months. In the letters forming a part of this pamphlet the statements on this phase of the question are quoted as follows: When poultry is to be frozen, the best stock in the market is generally selected; and if the same is killed and cooled properly, it can safely remain in cold storage for fully two years, if necessary, without becoming tainted and injurious to health. How- ever, it is seldom, or, I might say never, kept longer thaneight months. * * * Poultry that is dressed and cooled off properly before going into cold storage will come out in good wholesome condition any time, within at least one year, and it would take a very good expert to tell the difference between good fresh frozen poultry and fresh killed at time of eating. Another dealer states: Poultry with entrails in it can be kept in our modern freezers, where they shall be frozen and kept at a very low temperature, and be held in storage one, two, or three years, or even longer, without any detriment to the poultry itself, and on such poultry there is no possible chance of there being any cause for fear of ptomaine poisoning. Another warehouseman in a letter states: Poultry placed in a freezer under proper conditions and kept at the right temper- ature will keep an indefinite length of time and in healthful condition. Taking up the question of drawn and undrawn poultry, the attorneys for the warehousemen came to the conclusion that— The experience of men well versed in the trade as to the desirability of drawing poultry before storing, it is best shown by the letters printed herewith. From them it will be seen that drawing poultry before putting it in the freezer is impracticable. It will not keep as well drawn as undrawn. From the letters to which reference is made we find the following statements: We have experimented with poultry in every form. We have found that it is utterly impracticable to carry drawn poultry in the freezer. It is very similar to any other product in that so long as it remains an air-tight package it can be carried much better than when it is open to the atmosphere, which dries it, discolors, and generally injures the inside texture of the bird when exposed. Further, of the - markets which buy the frozen poultry from us there is not one that can or will accept it drawn; and although there is on the statute books in the State of Massachusetts a law demanding that all poultry marketed dressed shall be drawn, the same is a dead letter, and all our shipments to Boston and other Massachusetts cities are undrawn. Again, another dealer states: We are handling every year a small portion of storage poultry that is drawn, heads and feet off, and we usually have to sell this class of goods at from 1 to 3 cents below the price of the poultry that is undrawn with heads and feet on. I feel satisfied that if we had to draw our poultry and depend on our customers to sell the same to, we would have to quit the poultry business, as we certainly could not compete with other cities that are selling undrawn poultry. I might say that I do not know of any market in the United States where we would be able to sell drawn poultry, even at a discount of from 1 to 3 cents per pound less than the undrawn poultry. 11iQ PRELIMINARY COLD STORAGE STUDIES. In another letter occurs the following passage: A chicken when dead and undrawn is hermetically sealed, and you do not find the decomposition the greatest in the inside of the bird, but you do find it between the legs and the body and under the wings. In still another letter the warehouseman states: The result of having to draw poultry in the country for shipment into Chicago would cause the greater percentage oi the poultry to arrive at its destination in a more or less putrified condition, and such poultry put into storage, no matter under what conditions, even with the use of modern refrigeration, the inside of the poultry (the fact of its having been wet) will become moldy, either showing mold with black spots or with a woolly fungous growth. That opinion is not unanimous in regard to the advisability of retaining the entrails in dead animals until the time of cooking shall arrive is indicated by a number of public statements from various sources, certain of which are appended. VARIOUS OPINIONS ON DRAWN AND UNDRAWN POULTRY. Franklin G. Fay, of Sacramento, writing on the subject in the California State Journal of Medicine, 1904, Volume IV, page 66, states that— Resolutions condemning the use for human consumption of fish and poultry from which the viscera were not removed at the time of slaughter were passed by the Sacramento city board of health, and that such legislation, being before the Sacra- mento board of trustees, embodied in the form of an ordinance, was meeting with strenuous opposition on the part of the dealers, led by the large packers and cold- storage men. Doctor Fay says: Ii their immense interest in this matter is really in behalf of the public, we should have no difficulty in making a satisfactory adjustment, but as yet we have found no consumer outside of the trade who is opposed to legislation. * * * The reports oi the Canadian commissicners of agriculture show that the requirements of the English market demand that the intestines be removed. Among the very prominent opposers of the custom of marketing undrawn poultry is Doctor Cavana, of Oneida, N. Y., who makes the following statement in a paper entitled ‘‘Dangers in Undrawn Poultry and Game,” presented at the annual convention of the American Association of Railway Surgeons at Chicago, Ll., October 17, 1906: Bacterial cultures made from the breasts and legs from 100 undrawn fowls proved the presence and thorough permeation of the tissues in each specimen, with the vari- ous groups of intestinal germs, some tests showing no less than eleven distinct groups in one poultry specimen. Among the varieties of pathogenic bacteria identified was the Bacillus coli communis, the Staphylococcus pyogenes aureus, the Bacillus proteus vulgaris; and the Streptococcus pyogenes. Previous tests of intestinal matter taken irom the entrails of recently slaughtered fowls revealed the presence of these latter germs in great abundance in the intestinal canal of every fowl examined, and their discovery in the remote tissues of the undrawn cold-storage specimens proves un- APPENDIX. EVI ~ mistakably that their source was from the alimentary canal, and their permeation of the tissues the direct result of the retention of the intestinal tract and its contents in the sealed abdominal cavities of the poultry carcasses. Growers of poultry will sup- port us in the assertion that the ducks and hens of the barnyard are the scavengers of the farm. They are constantly picking over the soil, the farm garbage heaps, and other bacterial hotbeds, and no accretion or mass of decaying matter ever becomes too repulsive for poultry food, especially that of hens. In our bacteriological studies all of the cultures were made from the eatable tissues of the various specimens and with the most thorough aseptic precautions. The Bacillus coli communis was found in 100 per cent, or every specimen tested; the Bacillus proteus vulgaris in 6 per cent; the Staphylococcus pyogenes aureus in 20 per cent; and the Streptococcus pyogenes in 65 per cent of the 100 examinations. Probable infection before storage-—The cold storage plant owners of New York City inform us that their poultry stocks are collected from all parts of the country, even as far distant as the States of Texas, Louisiana, and Florida. After slaughter the feathers are removed and the carcasses packed in barrels without further dressing. The head, feet, and legs, as well as the craw of partially digested food, and the decom- posing livers, lungs, and intestines with their filthy contents, all combine to make valuable weight, and are therefore left in the sealed cavities of the fowls, forming conditions which force the general infection of the tissues by the flageilated, or rapidly swimming intestinal bacteria, which double their quantity and numbers every forty minutes, a single bacillus being capable of developing over forty-two billion germs in twenty-four hours. Their shipments are made by rail and steamship, and cover transit periods of several days before reaching the cold atmospheres of the storage warehouses. To determine the activity of these germs and the period required for their permea- tion of the tissues in the slaughtered undrawn fowl, we caused to be made a series of experiments, the results of which justify the belief that a great percentage of the infected poultry and game stock in storage became so infected before reaching the low temperature of the storage warehouses. . As a direct reply to the above-mentioned work of Doctor Cavana, Dr. Henry A. Higley, at the solicitation of the New York Poultry Dealers’ Association, made an investigation and presented the same at a hearing before the legislative committee at Albany, on behalf of the opposition to a bill introduced by Doctor Cavana providing that all poultry must be drawn within eighteen hours after killing. Among Doctor Higley’s conclusions are the following: 1. No matter how many of the bacteria which are concerned in this discussion there may be in the intestinal and thoracic cavities of dead undrawn poultry and game, they can not invade the edible portions so long as the temperature of such poultry and game is kept at 5° C. (41° F.) or below, because these bacteria do not grow at such a temperature or below. 2. Dead, undrawn poultry and game kept at a temperature (above 41° F.) which would allow the invasion of its edible portions with these bacteria mentioned from the intestinal tract would be subject to putrefaction changes because such a tempera- ture would be much more favorable for the growth of the bacteria which produce putrefaction, since they can grow at as low as 0° C. (32° F.). 3. Even if the bacteria invasion claimed by the supporters of this measure does take place, such bacteria can produce no poisonous substances because they are placed. under unfavorable conditions of growth so long as thé temperature of the fowls is kept anywhere near 5° C. (41° F.). 4. The longer dead poultry and game is kept frozen the less bacteria will it contain, because freezing temperatures gradually destroy bacteria. 1g PRELIMINARY COLD STORAGE STUDIES. And as a general conclusion to the whole matter he makes this statement: All bacteriological evidence conclusively proves that the edible portions of healthy, dead, undrawn poultry and game do not contain any bacteria, toxines, or ptomaines that are harmiul when eaten by man, so long as such poultry is kept free from putre- faction. That poultry that goes into cold storage in good bacterial condition comes out in exactly the same condition that it went in, so long as the temperature of the poultry is kept low enough to prevent the growth of putrefactive bacteria and, finally, that the longer poultry remains frozen the less bacteria does it contain. SUGGESTIVE LEGISLATIVE ACTION. MASSACHUSETTS ENACTMENTS. That the question of the advisability of storing fowls in an un- drawn condition is one of widespread interest is evidenced by the localities which have made this matter a subject of investigation and have finally presented it to their legislative bodies. For instance, in Massachusetts a bill was introduced relative to the sale and storage of poultry, in which it has recommended that— Whoever, himself or by his servant or agent, or as the servant or agent of any other person, firm, or corporation, sells, exposes for sale, exchanges or delivers, or has in his custody or possession with intent to sell, exchange, or deliver, any dead poultry from which the head, crop, if it contain food, and the entrails have not been removed, shall be punished by a fine of not less than five nor more than fifty dollars for each offense; but the provisions of this section shall not apply between the first day of April and the thirty-first day of October in the case of what is known commercially as ‘“‘Iced poultry,’ that is to say, poultry shipped in ice. Whoever, himself, or by his servant or agent, or as the servant or agent of any other person, firm, or corporation, places, or causes to be placed, or holds in cold storage for purposes of sale, any poultry from which the head, crop, if it contain food, and the entrails have not been removed, shall be punished by a fine of not less than twenty- five dollars for each day said poultry is held as aforesaid. For the purposes of this and the preceding section, the word ‘‘poultry” shall be deemed to mean ducks, geese, turkeys, fowls, and chickens. The result of the introduction of this bill was the passage of a resolution providing for a comparative investigation of drawn and undrawn poultry when shipped or stored. Dr. Charles Harrington, secretary of the board of health of Massachusetts, undertook to carry out the investigation which was recommended, and has sub- mitted a report in which he draws the following conclusions: 1. During cold storage at from 15 degrees below to 5 degrees above 0° Fahrenheit no chemical changes occur. This is shown by the absence in both the drawn and undrawn birds of ptomaines and decomposition products in general, and by negative reaction on the part of animals inoculated with extracts obtained from both kinds of material. 2. When removed from cold storage and exposed to ordinary temperatures, the condition of exposure being the same, the undrawn birds show better keeping quali- ties. 3. Freezing renders the muscular tissues more susceptible to bacterial invasion after they are thawed out. APPENDIX. Lie 4, The usual method of drawing poultry leads to heavy bacterial infection, which promotes more rapid decomposition than occurs in undrawn birds. 5. By ligature of the gullet below the crop, poultry can be completely drawn with- out any spilling of the intestinal contents, with consequent bacterial invasion of the abdominal cavity; and poultry so drawn would undoubtedly withstand decompo- sition and deterioration much longer than that which is undrawn. 6. The practice of depositing poultry in cold storage when in the beginning or advanced stages of decomposition, in order to save it, is dangerous to the health of the consumer, since when it is again withdrawn for sale its condition is unaltered. 7. Proper and adequate inspection of poultry as it enters cold storage is-desirable, and storage of material already in process of decomposition should be prohibited. 8. The practice of placing cold-storage poultry in cold water for a number of hours for the purpose of thawing causes heavy bacterial infection and consequent more rapid decomposition than occurs when thawing is allowed to proceed slowly at room temperature. Such treatment causes also a material increase in weight, by reason of absorption by the tissues of water, to the detriment of the purse of the purchaser, and hence is fraudulent. SACRAMENTO, CAL., ORDINANCE. The city of Sacramento has an ordinance relating to and regulating the sale of undrawn, slaughtered fish, game, and any animal to be used for food purposes within the limits of the city of Sacramento, and has prescribed a punishment for the violation of the provisions thereof. This ordinance forbids any person, firm, or corporation to sell, offer, or expose for sale any slaughtered animal used for food -purposes, refrigerated or otherwise, which has not been properly drawn and prepared by removing the viscera at the time of slaughter, and any person, firm, or corporation failing to follow the provisions of this ordinance is deemed guilty of a misdemeanor, and punished by a fine not exceeding $100 or by imprisonment in the city prison not exceeding fifty days, or by both such fine and imprisonment. This ordinance went into effect on and after the date of February 1, 1905. ENACTMENTS IN ILLINOIS. The Illinois State board of health has issued a pamphlet to the municipalities of the State of Illinois strongly recommending that there be enacted and enforced ordinances prohibiting the sale of fish, game, or any animal used for food which has not been properly drawn and cleaned at the time of slaughter. A paragraph in this pamphlet reads as follows: It is known to all physicians and physiologists that there are generated in the body of any animal poisons of the highest degree of toxicity. The intestines and other digestive organs contain at all times materials which have undergone putrefactive changes. If this material be permitted to remain in the body after death, the poisons generated may infiltrate the entire flesh, making it dangerous to the person who eats it. The body in which the viscera are permitted to remain undergoes decomposition much more rapidly than when such viscera have been removed. Decomposition is further hastened by leaving the blood in the vessels of the body. 49078—Bull. 115—08——8 114 PRELIMINARY COLD STORAGE STUDIES. Dr. E. N. Eckard, commissioner ot health of Peoria, Ill., caused to be instituted an examination of drawn and undrawn chickens, such as were found offered for sale in the markets of Peoria, coming from cold storage plants. On the basis of the reports aviemned to him by the bacteriologists in whose hands the practical investigation was placed, an ordinance was passed by the city council of Peoria, which reads as follows: Section 1. That it shall be unlawful for any person, firm, or corporation within the limits of the city of Peoria to sell, offer, or expose for sale, any animal, fowl, or game used for food purposes, refrigerated or otherwise, which has not been properly drawn and prepared by removing the viscera (bowels, entrails) at the time of slaughter. Sec. 2. Any person, firm, or corporation violating any of the provisions. of this ordinance shall be deemed guilty of a misdemeanor, and upon conviction thereof shall be punished by a fine not exceeding one hundred dollars. PROPOSED LEGISLATION IN NEW YORK. In the State of New York in January, 1907, there was offered the following amendment to Section 165 of the agricultural law, defining certain foods the sale of which is prohibited. If it consists of any slaughtered game, animal, poultry or fowl, unless the carcasses of such slaughtered game, animal, poultry, or fowl shall have been divested of its lung tissues, entire digestive and intestinal tracts, gall receptacle, craw, and gizzard lining, within twelve hours after its slaughter; and any slaughtered game, animal, poultry, or fowl found in any refrigerator, ice chest, cooler, storage apartment, or market, whether exposed for sale, or in stock, shall be presumed to have been slaugh- tered for a longer period than twelve hours. This amendment has been referred to under the discussion of the work of Doctor Cavana and Doctor Higley, respectively. PROPOSED LEGISLATION IN THE DISTRICT OF COLUMBIA. There was presented to the Fifty-ninth Congress on March 12,1906, a bill relating to the sale of poultry in the District of Columbia, which bill recommended that the shipping of any poultry into said District, intended for sale, refrigerated or otherwise, which has not been prop- erly drawn and prepared by removing the viscera at the time of slaughter shall be considered unlawful, a fine and imprisonment being provided as a punishment for the violation of this act. In the report of the committee to which this bill was referred there is a reeommenda- tion that it be passed. CANADIAN Ce OE ACT. An interesting contribution to the subject of cold storage is ane in the report of the dairy and cold storage commissioner of the Dominion of Canada for the year ending March 31, 1907. The Dominion of Canada has already secured the enactment of a law Wie, 5 * APPENDIX. sea Bes relating to the cold storage and regulations thereunder which are of interest in this connection. Certain portions of the act and regu- lations are as follows: THE COLD-STORAGE ACT. An act to encourage the establishment of cold-storage warehouses for the preservation of perish- able food products. His Majesty, by and with the advice and consent of the Senate and House of Com- mons of Canada, enacts as follows: 1. This act may be cited as the Cold Storage Act. 2. The Governor in Council may enter into contracts with any person for the con- struction, equipment, and maintenance in good and efficient working order of public cold-storage warehouses equipped with mechanical refrigeration, in Canada, and suitable for the preservation of all food products. 3. The location, plans, and specifications of every such warehouse, its equipment, and the amount to be expended thereon shall be subject to the approval of the Gov- ernor in Council. 4, The Governor in Council may, out of any moneys appropriated by Parliament for the purpose, grant toward the construction and equipment of any such warehouse a subsidy not exceeding in the whole 30 per cent of the amount expended or approved of in such construction and equipment, and payable in installments as follows: Upon the warehouse being completed and cold storage at suitable temperatures being pro- vided therein, all to the satisfaction of the Minister of Agriculture, asum not exceeding 15 per cent of the amount so expended; and at the end of the first year thereafter 7 per cent of the said amount; at the end of the second year thereafter, 4 per cent of the said amount, and at the end of each of the two next succeeding years, 2 per cent of the said amount: Provided, The warehouse is maintained and operated to the satis- faction of the Minister of Agriculture. 5. The Minister of Agriculture may refuse to pay any part of the said subsidy if, in his opinion, the operation of the warehouse has not been of such a character as to pro- vide for the proper preservation of such products as may be stored therein. 6. The Minister of Agriculture may order, and cause to be maintained, an inspection and supervision of the sanitary conditions, maintenance, and operation of such ware- houses, and may regulate and control the temperatures to be maintained therein in accordance with the regulations to be made as hereinafter provided. 7. The rates and tolls to be charged for storage in such warehouses shall be subject to the approval of the Governor in Council. 8. For the effective carrying out of the provisions of this Act the Minister of Agri- culture may appoint inspectors, who shall have access to all parts of such warehouses at all times. 9. The Governor in Council may make such regulations as he considers necessary in order to secure the efficient enforcement and operation of this Act; and he may by such regulations impose penalties not exceeding fifty dollars on any person offending against them; and the regulations so made shall be in force from the date of their pub- lication in the Canada Gazette, or from such other date as is specified in the proclama- tion in that behalf. Among the regulations for the enforcement and operation of this act are the following: 2. No applications shall be considered for any cold-storage warehouses except those equipped with mechanical refrigeration, nor for any place where any such cold storage already exists or where the proposed cold storage would compete nea with other establishments of the same class. * * * 116 PRELIMINARY COLD STORAGE STUDIES. : 4. The owners of cold-storage warehouses, in order to secure the subsidy, will be required to maintain the following temperatures therein for the preservation of the various products mentioned: Temperature (° F.). Kinds of produce. SSS Minimum. | Maximum. Apples:and other iruitss22%. S28 $< a ase ae St ee ee ae eee eae 32 36 PSU GEOL Sach sre Se Se a ees ed Beer ee ae 20 Cheese 2 22 soe 2 SS a ee eee er Ee 35 40 Eggs, meats, and dressed poultry..------ ee ee ee eds te eee AR Ve ee et ea ae 30 34 BaGon and: Wams'ssai co a ee re ce 40- 45 ish Grozen) "LE ikce ee see eee ah Sonora eee ee fos Sone ee eee |e abiscemereree 20 Meats: poultry, amd camer ro Zen) eee se me ere |r Stes eres 20 Vesetales sos. Se ape a oe a eee Re ame ne 34 38 5. Nothing in these regulations shall prevent owners of subsidized cold-storage warehouses from entering into special contracts with customers for the maintenance of temperatures other than those herein specified. * * * 7. The owners of cold-storage warehouses, to which the subsidy or any part thereof has been paid, may be required to make an annual report to the Minister of Agriculture in such form as may be prescribed. The subject of cold storage is of great importance in Canada, par- ticularly in relation to the export of food products. The rapid increase of population in the western part of the Dominion, where the production of perishable products of the farm is not large enough to supply the local needs, has, however, developed a great internal trade in cold-storage products. It is said by the Commissioner on page 156 of his report: Cold storage enables the dairyman to store his surplus butter and cheese during the active season of production ana to dispose of it during the off season, when manu- facture has almost, if not entirely, ceased. Prices are equalized, and the consumer is supplied with an article in better condition throughout the whole year than would otherwise be possible. Not only is the business of the producer enormously increased, but the commerce of the country derives a corresponding benefit. The Dominion of Canada has encouraged the industry of cold storage by offering certain subsidies, as is provided for in the bill preceding. The dairy and cold storage commissioner says in regard to this matter: The policy of giving financial assistance toward the erection of public cold-storage warehouses in Canada was adopted by the Government during the session of 1906-7 by the introduction of the Cold Storage Bill, entitled an ‘‘Act to encourage the estab- lishment of cold-storage warehouses for the preservation of perishable food priducie. Parliament approved of the measure, and it became law without delay. The ee of trade in certain localities have made cold storage an absolute necessity, and in such places the revenue is sure enough to make the investment a fairly safe one, so that it needed no special inducement to secure the capitai required to provide the necessary facilities. While it may, at first glance, seem a little unfair to assist the new enterprises, even if noncompetitive, it must be borne in mind that the existing cold-storage warehouses occupy the choice focrnees and for that reason some inducement seems to be necessary: if similar facilities are to be provided im other localities. APPENDIX. TEs % There is a large quantity of perishable produce handled in this country without cold storage, the value and stability of which would be much more improved by its use. Where it is possible to get along without cold storage, even if the results are unsatis- factory, the question is not studied so closely, and the improved facilities come more slowly than is the case where the necessities are greater. It is hoped that the attention which has been drawn to the subject by the adoption of the principles involved in the Cold Storage Act, and the discussion which will naturally arise thereon, will have an important educational influence in the direction of creating a more general appreciation of the advantages of cold storage and a greater demand for such facilities. It is believed also that this process of education will result in bringing more business to existing cold- storage warehouses. It will be seen by an inspection of the quotations made from the report and comments thereon that the object of the regulation of cold storage in Canada is more for the development of commerce than it is to study the effects of cold storage upon the wholesomeness of the products. Nevertheless, sanitary provisions are included in the act in section 5, where the subsidy which is offered may be refused if precautions for the proper preservation of the products have not been observed, and also in section 6, where a sanitary inspection and supervision of the cold-storage plants are provided for. The Dominion of Canada, it appears, has taken a more active interest officially in cold storage than any other country so far as is known, since they have recognized it by an act of Parliament and provided to a certain extent for its regulation. O